Development of Non-Overlapping Multiplex ELISA Arrays for the Quantitative Measurement of 400 Human and 200 Mouse Proteins in Parallel

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Development of non-overlapping multiplex ELISA arrays for the quantitative measurement of 400 human and 200 mouse proteins in parallel

Yingqing Mao1, Zhiqiang Lv1, Haw-Han Yen1, Yanni Sun1 and Ruo-Pan Huang1,2,3

1RayBiotech, Inc., 3607 Parkway Lane, Suite 100, Norcross, GA 30092, USA; 2RayBiotech Inc., Guangzhou, 510600 China; 3South China Biochip Research Center, Guangzhou, China 510630

Web: www.raybiotech.com Email: [email protected] Phone: 770-729-2992

Abstract Platform & Principal Sensitivity & X-reactivity Highlights

Accurate detection of multiple cytokines in complex biological samples is essential to progress in immunological research. However, development of Human receptor array • Proven sandwich ELISA detection high-density multiplex sandwich-based ELISA panels is hampered by 60,000 50,000 Control Array sensitivity test nonspecific cross-reactivity between and among target-specific reagents 40,000 Std7 (Left): One set of 3x • High throughput multiplex array format (i.e., capture antibodies, detection antibodies, and/or antigens); this is 30,000 Std6 Std5 diluted standards were 20,000 particularly challenging for bead-based assays, in which all three are free to Std4 run on human receptor 10,000 • Quadruplicate data for each target per run interact in solution. To overcome this bottleneck, we tested for nonspecific Std3

0 Std2 array.

3 1

1 2

- - -

Fas

Dtk

1 RI 1

17R 21R

1BB

DR6

2 Rg 2

1 R4 1

GITR

- - -

- Std1

uPAR CD14 CD30

MICB

MICA

RAGE

10 Rb 10 Flt

cross-reactivity among reagents used on our quantitative antibody arrays (in TIM

ErbB3

BCMA

HVEM

LIMPII

- IL IL

XEDAR

LYVE

CD40L

ALCAM

ICAM

Selectin Selectin

Fluor dye (cy3 equivalent) IL

VCAM

PDGF Rb

Endoglin

- TRAIL R3 TRAIL

NRG1 • ELISA sensitivity with wider dynamic range

PECAM

Trappin

E CEACAM which capture antibodies are affixed to a glass slide). Combinations of Lipocalin Biotin-Streptavidin complex reagents exhibiting nonspecific interactions were separated on discrete B Detect antibody • High specificity, accuracy & reproducibility arrays. Using this divide-and-conquer strategy, we developed and validated Cytokine 25,000 10 human and 5 mouse non-overlapping 40-plex arrays for parallel, Capture antibody Array X-reactivity test 20,000 • Less sample, time, and cost

quantitative detection of 400 human and 200 mouse proteins respectively. To Glass Slide Support (Right): Individual DAB 15,000

our best knowledge, this is the highest density multiplex ELISA developed was tested in the 10,000 TRAIL R3 NRG-b1 • Suitable for diverse sample types 5,000 Lipocalin-2 to date. Such a system hold a great promise in determining key factors and Configuration of array platform and illustration of sandwich presence of mixed IL-10 Rb 0 GITR mechanisms in immune cell physiology and pathology, in identifying novel ELISA principal. One standard glass slide is spotted with 16 wells of antigens (10 ng/ml Endoglin identical cytokine antibody arrays. The sandwich ELISA detection CD30 • Software supports one step data computation each) with all CAB. 4-1BB drug targets, and in screening for putative biomarkers from patients' principal is depicted below after the consequent reaction of sample, samples. biotinylated detection antibody, and Streptavidin labeled fluor dye.

Strategy Array vs. ELISA Accuracy & Precision Sample Test

Step 1: ELISA confirmation of cytokine antibody IL-6 Detection Dynamic Range Example of Levey Jennings (pg/ml) LOD Normal Serum (n=31) pairs; System optimization in array platform. in Quantibody and ELISA Quadruplicate data IL-1a 5 6 ± 2 106 Chart for IL-6 in 14 runs Coefficient of Variation < IL-1b 1 1 ± 0 IL-1ra 59 222 ± 60 150.3 105 (n=8124) LOD IL-2 5 27 ± 5 IL-4 4 7 ± 2 Step 2: Print capture antibody(CAB) arrays, and 4 35 17% 10 140.3 UCL 139.53 IL-5 11 82 ± 15 test individual detection antibody (DAB) in the IL-6 7 42 ± 7 3 10 IL-6sR 7 755 ± 25 presence of mixed antigens at 10 ng/ml each. ELISA 30 < Quantibody 130.3 IL-7 11 50 ± 6 102 > 25 LOQ IL-8 1 13 ± 2 IL-10 3 7 ± 1 101 120.3 LOQ 16% IL-11 34 39 ± 11

Step 3: Do inter-array signal normalization, and 0 20 IL-12p40 18 30 ± 4 10 CL 109.87 67% create cross reactivity table (DAB vs. CAB). 110.3 IL-12p70 1 0 ± 0 10-1 15 IL-13 3 16 ± 3 IL-15 17 158 ± 33 Signal Intensity (Quantibody) or OD (ELISA) OD or (Quantibody) Intensity Signal 100.3

10-2 IL-16 16 69 ± 14 6 Concentration Concentration 6 (pg/ml)

0.1 1 10 100 1000 10000 - 10 IL-17 18 2 ± 0 IL

Step 4: Divide cytokines into different subgroups IL-6 Concentration (pg/ml) 90.3 MCP-1 7 180 ± 23 Percentage of Assays of Percentage based upon the serum abundance, functionality 5 MCSF 3 1 ± 0 LCL 80.21 etc. and generate multiple 40-plex arrays. 80.3 Comparison of array and ELISA dynamic range and detection sensitivity. The 0 <5% 5-10% 10-15% 15-20% >20% dynamic range of IL-6 in array and ELISA (Left) and the correlation of IL-6 detection 70.3 Cytokine quantification in normal human sera (n=31). CV Range Step 5:Test individual array sensitivity, cross by both methods in 40 human culture media samples (Right). The same IL-6 pair of Limit of Detection (LOD): 3 standard deviation above negative control. reactivity, and reproducibility. Real sample test. antibodies together with the same antigen resource were used in both systems. Limit of Quantification (LOQ) = 10 standard deviation above negative control.

Array Development Multiplex ELISA Arrays List of Target Proteins Conclusions

Human targets Mouse targets Individual cross reactivity test Cross Reactivity Database 70000 IL-17 1. Here we developed and validated multiple non- # Cytokine Gene Symbol Uniprot Number HUGO Approved Name # Cytokine Gene Symbol Uniprot Number HUGO Approved Name # Cytokine Gene Symbol Uniprot Number HUGO Approved Name # Cytokine Gene Symbol Uniprot Number HUGO Approved Name # Cytokine In Array Gene Symbol # Cytokine In Array Gene Symbol 60000 1 2B4 CD244 Q9BZW8 CD244 molecule, natural killer cell receptor 2B4 101 Endostatin COL18A1 P39060 collagen, type XVIII, alpha 1 201 IL-1F6 IL36A Q9UHA7 interleukin 36, alpha 301 P-Cadherin CDH3 P22223 cadherin 3, type 1, P-cadherin (placental) 1 4-1BB mCYT6 TNFRSF9 101 IL-21 mCYT5 IL21 5PL Curve Signal Intensity 2 4-1BB TNFRSF9 Q07011 tumor necrosis factor receptor superfamily, member 9 102 Eotaxin-1 CCL11 P51671 chemokine (C-C motif) ligand 11 202 IL-1F7 IL37 Q9NZH6 interleukin 37 302 PD-1 PDCD1 Q15116 programmed cell death 1 2 6Ckine mCYT8 CCL21 102 IL-22 mCYT8 IL22 Array Cross Reactivity Check 3 6Ckine CCL21 O00585 chemokine (C-C motif) ligand 21 103 Eotaxin-2 Ccl24 O00175 chemokine (C-C motif) ligand 24 203 IL-1F8 IL36B Q9NZH7 interleukin 36, beta 303 PD-ECGF TYMP P19971 thymidine phosphorylase 3 ACE mCYT6 ACE 103 IL-23 mCYT4 IL23A CEA 4 A2M A2M P01023 alpha-2-macroglobulin 104 Eotaxin-3 CCL26 Q9Y258 chemokine (C-C motif) ligand 26 204 IL-1F9 IL36G Q9NZH8 interleukin 36, gamma 304 PDGF R alpha PDGFRA P16234 platelet-derived growth factor receptor, alpha polypeptide 4 Activin A mCYT8 INHBA 104 IL-28 mCYT4 IFNL2 50000 5 ACE-2 ACE2 Q9BYF1 angiotensin I converting enzyme 2 105 EpCAM EPCAM P16422 epithelial cell adhesion molecule 205 IL-1R3 IL1RAP Q9NPH3 interleukin 1 receptor accessory protein 305 PDGF Rb PDGFRB P09619 platelet-derived growth factor receptor, beta polypeptide 2,000 5 ADAMTS1 mCYT8 ADAMTS1 105 IL-3 mCYT5 IL3 3500 6 Activin A INHBA P08476 inhibin, beta A 106 Epo R EPOR P19235 erythropoietin receptor 206 IL-1R5 Il18r1 Q13478 interleukin 18 receptor 1 306 PDGF-AA PDGFA P04085 platelet-derived growth factor alpha polypeptide 6 Adiponectin mCYT8 ADIPOQ 106 IL-3 Rb mCYT6 Csf2rb 7 ADAM-9 ADAM9 Q13443 ADAM metallopeptidase domain 9 107 ErbB2 ErbB2 P04626 v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 2 207 IL-1ra IL1RN P18510 interleukin 1 receptor antagonist 307 PDGF-AB PDGFB P01127 platelet-derived growth factor beta polypeptide overlapping multiplex ELISA arrays for quantitative 7 ALK-1 mCYT6 SLPI 107 IL-33 mCYT7 IL33 8 Adiponectin ADIPOQ Q15848 adiponectin, C1Q and collagen domain containing 108 ErbB3 ErbB3 P21860 v-erb-b2 avian erythroblastic leukemia viral oncogene homolog 3 208 IL-2 IL2 P60568 interleukin 2 308 PDGF-BB PDGFB P01127 platelet-derived growth factor beta polypeptide 8 ANG-3 mCYT8 ANGPTL1 108 IL-4 mCYT5 IL4 3000 40000 9 Adipsin CFD P00746 complement factor D (adipsin) 109 E-Selectin SELE P16581 selectin E 209 IL-2 Ra IL2RA P01589 interleukin 2 receptor, alpha 309 PECAM-1 PECAM1 P16284 platelet/endothelial cell adhesion molecule 1 9 ANGPTL3 mCYT8 ANGPTL3 109 IL-5 mCYT5 IL5 10 aFGF FGF1 P05230 fibroblast growth factor 1 (acidic) 110 FABP2 FABP2 P12104 fatty acid binding protein 2, intestinal 210 IL-2 Rb IL2RA P01589 interleukin 2 receptor, alpha 310 Pepsinogen I PGA3 P0DJD8 pepsinogen 3, group I (pepsinogen A) 10 AR mCYT4 AREG 110 IL-6 mCYT5 IL6 1,500 11 AFP AFP P02771 alpha-fetoprotein 111 FAP FAP Q12884 fibroblast activation protein, alpha 211 IL-2 Rg IL2RG P31785 interleukin 2 receptor, gamma 311 Pepsinogen II PGC P20142 progastricsin (pepsinogen C) 11 Artemin mCYT8 ARTN 111 IL-7 mCYT5 IL7 2500 12 AgRP AgRP O00253 agouti related protein homolog (mouse) 112 FAS Fas P25445 Fas cell surface death receptor 212 IL-20 IL20 Q9NYY1 interleukin 20 312 Periostin POSTN Q15063 periostin, osteoblast specific factor 12 Axl mCYT4 Axl 112 IL-7 Rα mCYT7 IL7R 30000 13 Albumin ALB P02768 albumin 113 FAS L FASLG P48023 Fas ligand (TNF superfamily, member 6) 213 IL-21 IL21 Q9HBE4 interleukin 21 313 Persephin PSPN O60542 persephin 13 B7-1 mCYT7 CD80 113 IL-9 mCYT6 IL9 14 ALCAM ALCAM Q13740 activated leukocyte cell adhesion molecule 114 Fcr RIIBC FCGR2B P31994 Fc fragment of IgG, low affinity IIb, receptor (CD32) 214 IL-21R IL21R Q9HBE5 interleukin 21 receptor 314 PF4 CXCL14 O95715 chemokine (C-X-C motif) ligand 14 14 BAFF R mCYT7 TNFRSF13C 114 I-TAC mCYT4 CXCL11 measurement of 400 human and 200 mouse proteins. 2000 15 AMICA AMICA1 Q86YT9 adhesion molecule, interacts with CXADR antigen 1 115 Ferritin FTH1 P02794 ferritin, heavy polypeptide 1 215 IL-23 IL23A Q9NPF7 interleukin 23, alpha subunit p19 315 PGRP-S PGLYRP1 O75594 peptidoglycan recognition protein 1 15 bFGF mCYT5 FGF2 115 JAM-A mCYT6 F11R 1,000 16 ANG ANG P03950 angiogenin, ribonuclease, RNase A family, 5 116 Fetuin A AHSG P02765 alpha-2-HS-glycoprotein 216 IL-24 IL24 Q13007 interleukin 24 316 PIGF PGF P49763 placental growth factor 16 BLC mCYT5 CXCL13 116 KC mCYT5 CXCL1 20000 17 ANG-1 ANGPT1 Q15389 angiopoietin 1 117 FGF-19 FGF19 O95750 fibroblast growth factor 19 217 IL-27 IL27 Q8NEV9 interleukin 27 317 Procalcitonin CALCA P01258 calcitonin-related polypeptide alpha 17 BTC mCYT7 BTC 117 Kremen-1 mCYT7 KREMEN1 1500 BKG) - (Signal Intensity 18 ANG-2 ANGPT2 O15123 angiopoietin 2 118 FGF-4 FGF4 P08620 fibroblast growth factor 4 218 IL-28A IFNL2 Q8IZJ0 interferon, lambda 2 318 Prolactin PRL P01236 prolactin 18 C5a mCYT7 C5 118 Leptin mCYT5 LEP 19 ANG-4 ANGPT4 Q9Y264 angiopoietin 4 119 FGF-6 FGF6 P10767 fibroblast growth factor 6 219 IL-29 IFNL1 Q8IU54 interferon, lambda 1 319 Prostasin PRSS8 Q16651 protease, serine, 8 19 CCL28 mCYT8 CCL28 119 Leptin R mCYT6 LEPR Signal IntensitySignal 20 Angiostatin PLG P00747 plasminogen 120 FGF-7 FGF7 P21781 fibroblast growth factor 7 220 IL-3 IL3 P08700 interleukin 3 (colony-stimulating factor, multiple) 320 PSA KLK3 P07288 kallikrein-related peptidase 3 20 CCL6 mCYT7 CCL6 120 Limitin mCYT7 IFNZ 1000 10000 500 21 ANGPTL3 ANGPTL3 Q9Y5C1 angiopoietin-like 3 121 FGF-9 FGF9 P31371 fibroblast growth factor 9 221 IL-31 IL31 Q6EBC2 interleukin 31 321 PSMA FOLH1 Q04609 folate hydrolase (prostate-specific membrane antigen) 1 21 CD27 mCYT6 CD27 121 Lipocalin-2 mCYT7 LCN2 22 ANGPTL4 ANGPTL4 Q9BY76 angiopoietin-like 4 122 Fibrinogen FGA P02671 fibrinogen alpha chain 222 IL-33 IL33 O95760 interleukin 33 322 PTH PTH P01270 parathyroid hormone 22 CD27L mCYT4 CD27LG 122 LIX mCYT5 Cxcl5 23 ApoA1 APOA1 P02647 apolipoprotein A-I 123 FLRG FSTL3 O95633 follistatin-like 3 (secreted glycoprotein) 223 IL-34 IL34 Q6ZMJ4 interleukin 34 323 RAGE MOK Q9UQ07 MOK protein kinase 23 CD30L mCYT5 CD30LG 123 LOX-1 mCYT7 OLR1 500 24 ApoC1 ApoC1 P02654 apolipoprotein C-I 124 Flt-3L FLT3LG P49771 fms-related tyrosine kinase 3 ligand 224 IL-4 IL4 P05112 interleukin 4 324 RANK TNFRSF11A Q9Y6Q6 tumor necrosis factor receptor superfamily, member 11a, NFKB activator 24 CD30T mCYT4 CD30 124 L-Selectin mCYT6 SELL 25 ApoC2 ApoC2 P02655 apolipoprotein C-II 125 Follistatin FST P19883 follistatin 225 IL-5 IL5 P05113 interleukin 5 (colony-stimulating factor, eosinophil) 325 RANTES CCL5 P13501 chemokine (C-C motif) ligand 5 25 CD36 mCYT8 CD36 125 Lymphotactin mCYT6 XCL1 2. These proteins cover the most analyzed and biologically 0 0 26 ApoC3 ApoC3 P02656 apolipoprotein C-III 126 FOLR1 FOLR1 P15328 folate receptor 1 (adult) 226 IL-5 Ra IL5RA Q01344 interleukin 5 receptor, alpha 326 RAP LRPAP1 P30533 low density lipoprotein receptor-related protein associated protein 1 26 CD40 mCYT4 CD40 126 MadCAM-1 mCYT6 MADCAM1 0 0 100 200 300 400 500 600 700 800 900 27 ApoE ApoE P02649 apolipoprotein E 127 FSH FSHB P01225 follicle stimulating hormone, beta polypeptide 227 IL-6 IL6 P05231 interleukin 6 (interferon, beta 2) 327 RBP4 RBP4 P02753 retinol binding protein 4, plasma 27 CD40L mCYT6 CD40LG 127 Marapsin mCYT7 PRSS27

0 1 10 100 1,000 10,000 28 ApoH ApoH P02749 apolipoprotein H (beta-2-glycoprotein I) 128 Furin Furin P09958 furin (paired basic amino acid cleaving enzyme) 228 IL-6R IL6R P08887 interleukin 6 receptor 328 Resistin RETN Q9HD89 resistin 28 CD48 mCYT7 CD48 128 MBL-2 mCYT7 MBL2

78 27

16 29 APRIL TNFSF13 O75888 tumor necrosis factor (ligand) superfamily, member 13 129 Galectin-3 LGALS3 P17931 lectin, galactoside-binding, soluble, 3 229 IL-7 IL7 P13232 interleukin 7 329 SAA SAA1 P0DJI8 serum amyloid A1

b2 10

BB 29 CD6 mCYT7 CD6 129 MCP-1 mCYT5 CCL2

- -

BLC

BTC MIF

CEA

2 Rb 2

MSP

NOV B7

OSM 30 AR AREG P15514 amphiregulin 130 Galectin-7 LGALS7 P47929 lectin, galactoside-binding, soluble, 7 230 IL-8 IL8 P10145 interleukin 8 330 SCF KITLG P21583 KIT ligand Gas1

aFGF 30 Chemerin mCYT7 RARRES2 130 MCP-5 mCYT5 Ccl12

IL IL

100b

DcR3 TACE

TNFb CD14

10 Ra 10 Cytokine Concentration (pg/ml)

Flt

18 Rb 18

ErbB3

LDL R

- HCC

- Cytokine Concentration (pg/ml)

ANG

Insulin

MCP IL

S 31 Artemin ARTN Q5T4W7 artemin 131 Gas1 GAS1 P54826 growth arrest-specific 1 231 IL-9 IL9 P15248 interleukin 9 331 SCF R KIT P10721 v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog TGF

TIMP 31 Chordin mCYT8 CHRD 131 M-CSF mCYT5 CSF1

sgp130

WISP

CRTAM

ICAM ENA

1/OLR1

TREM

IGFBP

MMP

PDGF Prolactin Cadherin 32 Axl Axl P30530 AXL receptor tyrosine kinase 132 GASP-1 GPRASP1 Q5JY77 G protein-coupled receptor associated sorting protein 1 232 Insulin INS P01308 insulin 332 SDF-1a CXCL12 P48061 chemokine (C-X-C motif) ligand 12 32 Clusterin mCYT7 CLU 132 MDC mCYT4 CCL22 - important secreted molecules such as cytokines,

33 B2M B2M P61769 beta-2-microglobulin 133 GASP-2 GPRASP2 Q96D09 G protein-coupled receptor associated sorting protein 2 233 IP-10 CXCL10 P02778 chemokine (C-X-C motif) ligand 10 333 SDF-1b CXCL12 P48061 chemokine (C-X-C motif) ligand 12 33 CRP mCYT8 CRP 133 Meteorin mCYT7 METRN

VE LOX DPPIV/CD26 34 B7-1 CD80 P33681 CD80 molecule 134 GCP-2 CXCL6 P80162 chemokine (C-X-C motif) ligand 6 234 I-TAC CXCL11 O14625 chemokine (C-X-C motif) ligand 11 334 Serpin A4 SERPINA4 P29622 serpin peptidase inhibitor, clade A, member 4 34 CT-1 mCYT6 CTF1 134 MFG-E8 mCYT6 Mfge8 Fetuin A/AHSGFetuin 35 BAFF TNFSF13B Q9Y275 tumor necrosis factor (ligand) superfamily, member 13b 135 G-CSF CSF3 P09919 colony stimulating factor 3 (granulocyte) 235 Kallikrein 14 KLK14 Q9P0G3 kallikrein-related peptidase 14 335 Serpin F1 SERPINF1 P36955 serpin peptidase inhibitor, clade F, member 1 35 CTLA-4 mCYT6 Ctla4 135 MIG mCYT5 CXCL9 36 BCAM BCAM P50895 basal cell adhesion molecule (Lutheran blood group) 136 G-CSF R CSF3R Q99062 colony stimulating factor 3 receptor (granulocyte) 236 Lactoferrin LTF P02788 lactotransferrin 336 sFRP-3 FRZB Q92765 frizzled-related protein 36 CXCL15 mCYT7 CXCL15 136 MIP-1a mCYT5 CCL3

37 BCMA TNFRSF17 Q02223 tumor necrosis factor receptor superfamily, member 17 137 GDF-15 GDF15 Q99988 growth differentiation factor 15 237 LAG-3 LAG3 P18627 lymphocyte-activation gene 3 337 sgp130 Il6st P40189 interleukin 6 signal transducer (gp130, oncostatin M receptor) 37 CXCL16 mCYT4 CXCL16 137 MIP-1b mCYT8 CCL4 2B4/CD244/SLAMF4 38 BDNF BDNF P23560 brain-derived neurotrophic factor 138 GDNF GDNF P39905 glial cell derived neurotrophic factor 238 LAP/TGF beta 1 TGFB1 P01137 transforming growth factor, beta 1 338 Shh N SHH Q15465 sonic hedgehog 38 Cystatin C mCYT7 CST3 138 MIP-1g mCYT5 Ccl9 39 bFGF FGF2 P09038 fibroblast growth factor 2 (basic) 139 GH GH1 P01241 growth hormone 1 239 Layilin LAYN Q6UX15 layilin 339 SIGIRR SIGIRR Q6IA17 single immunoglobulin and toll-interleukin 1 receptor (TIR) domain 39 DAN mCYT7 PARN 139 MIP-2 mCYT4 Cxcl2 Multiplex standard curves 40 bIG-H3 TGFBI Q15582 transforming growth factor, beta-induced, 68kDa 140 GITR TNFRSF18 Q9Y5U5 tumor necrosis factor receptor superfamily, member 18 240 LDL R LDLR P01130 low density lipoprotein receptor 340 Siglec-5 SIGLEC5 O15389 sialic acid binding Ig-like lectin 5 40 Decorin mCYT6 DCN 140 MIP-3a mCYT4 CCL20 chemokines, growth factors, soluble receptors, and other 41 BLC CXCL13 O43927 chemokine (C-X-C motif) ligand 13 141 GITR Ligand TNFSF18 Q9UNG2 tumor necrosis factor (ligand) superfamily, member 18 241 Legumain LGMN Q99538 legumain 341 Siglec-9 SIGLEC9 Q9Y336 sialic acid binding Ig-like lectin 9 41 Dkk-1 mCYT6 DKK1 141 MIP-3b mCYT6 CCL19 42 BMP-2 BMP2 P12643 bone morphogenetic protein 2 142 GM-CSF CSF2 P04141 colony stimulating factor 2 (granulocyte-macrophage) 242 Leptin LEP P41159 leptin 342 SOST SOST Q9BQB4 sclerostin 42 DLL4 mCYT7 DLL4 142 MMP-10 mCYT8 MMP10 43 BMP-4 BMP4 P12644 bone morphogenetic protein 4 143 GPBB PYGB P11216 phosphorylase, glycogen; brain 243 Leptin R LEPR P48357 leptin receptor 343 ST2 IL1RL1 Q01638 interleukin 1 receptor-like 1 43 Dtk mCYT6 TYRO3 143 MMP-2 mCYT8 MMP2 44 BMP-5 BMP5 P22003 bone morphogenetic protein 5 144 GRO CXCL1 P09341 chemokine (C-X-C motif) ligand 244 LIF LIF P15018 leukemia inhibitory factor 344 Syndecan-1 SDC1 P18827 syndecan 1 44 E-Cadherin mCYT8 CDH1 144 MMP-3 mCYT8 MMP3 45 BMP-6 BMP6 P22004 bone morphogenetic protein 6 145 GROa CXCL1 P09341 melanoma growth stimulating activity, alpha 245 LIGHT TNFSF14 O43557 tumor necrosis factor (ligand) superfamily, member 14 345 TACE ADAM17 P78536 ADAM metallopeptidase domain 17 45 EDAR mCYT7 EDAR 145 Neprilysin mCYT6 MME 46 BMP-7 BMP7 P18075 bone morphogenetic protein 7 146 HAI-2 SPINT2 O43291 serine peptidase inhibitor, Kunitz type, 2 246 LIMPII SCARB2 Q14108 scavenger receptor class B, member 2 346 TACI TNFRSF13B O14836 tumor necrosis factor receptor superfamily, member 13B 46 EGF mCYT4 EGF 146 Nope mCYT7 IGDCC4 47 BMP-9 GDF2 Q9UK05 growth differentiation factor 2 147 HB-EGF HBEGF Q99075 heparin-binding EGF-like growth factor 247 Lipocalin-2 LCN2 P80188 lipocalin 2 347 TARC CCL17 Q92583 chemokine (C-C motif) ligand 17 47 Endocan mCYT7 ESM1 147 NOV mCYT7 NOV regulatory factors. 48 b-NGF NGF P01138 nerve growth factor (beta polypeptide) 148 HCC-1 CCL14 Q16627 chemokine (C-C motif) ligand 14 248 LOX-1 OLR1 P78380 oxidized low density lipoprotein (lectin-like) receptor 1 348 TECK CCL25 O15444 chemokine (C-C motif) ligand 25 48 Endoglin mCYT6 ENG 148 OPG mCYT4 TNFRSF11B 49 BTC BTC P35070 betacellulin 149 HCC-4 CCL16 O15467 chemokine (C-C motif) ligand 16 249 L-Selectin SELL P14151 selectin L 349 TFPI TFPI P10646 tissue factor pathway inhibitor 49 Eotaxin mCYT5 CCL11 149 OPN mCYT4 SPP1 50 C5a C5 P01031 complement component 5 150 hCGb CGB P01233 chorionic gonadotropin, beta polypeptide 250 Lymphotactin XCL1 P47992 chemokine (C motif) ligand 1 350 TGFa TGFA P01135 transforming growth factor, alpha 50 Eotaxin-2 mCYT5 Ccl24 150 Osteoactivin mCYT7 GPNMB 51 CA125 MUC16 Q8WXI7 mucin 16, cell surface associated 151 Hemoglobin HBA1 P69905 hemoglobin, alpha 1 251 LYVE-1 LYVE1 Q9Y5Y7 lymphatic vessel endothelial hyaluronan receptor 1 351 TGFb RIII TGFBR3 Q03167 transforming growth factor, beta receptor III 51 Epigen mCYT8 EPGN 151 OX40 Ligand mCYT7 TNFSF4 52 CA15-3 MUC1 P15941 mucin 1, cell surface associated 152 HGF HGF P14210 hepatocyte growth factor (hepapoietin A; scatter factor) 252 Marapsin PRSS27 Q9BQR3 protease, serine 27 352 TGFb1 TGFB1 P01137 transforming growth factor, beta 1 52 Epiregulin mCYT8 EREG 152 P-Cadherin mCYT7 CDH3 53 CA199 #N/A #N/A Cancer antigen 19-9 153 HGF R MET P08581 met proto-oncogene 253 MBL MBL2 P11226 mannose-binding lectin (protein C) 2, soluble 353 TGFb2 TGFB2 P61812 transforming growth factor, beta 2 53 E-selectin mCYT4 SELE 153 PDGF-AA mCYT8 PDGFA 54 CA9 CA9 Q16790 carbonic anhydrase IX 154 HTRA2 HTRA2 O43464 HtrA serine peptidase 2 254 MCF R CSF1R P07333 colony stimulating factor 1 receptor 354 TGFb3 TGFB3 P10600 transforming growth factor, beta 3 54 Fas mCYT8 Fas 154 Pentraxin 3 mCYT6 PTX3 55 Calbindin D CALB1 P05937 calbindin 1, 28kDa 155 HVEM TNFRSF14 Q92956 tumor necrosis factor receptor superfamily, member 14 255 MCP-1 CCL2 P13500 chemokine (C-C motif) ligand 2 355 Thrombomodulin THBD P07204 thrombomodulin 55 Fas L mCYT5 FASLG 155 Periostin mCYT7 POSTN 56 Cathepsin S CTSS P25774 cathepsin S 156 I-309 CCL1 P22362 chemokine (C-C motif) ligand 1 256 MCP-2 CCL8 P80075 chemokine (C-C motif) ligand 8 356 Thyroglobulin TG P01266 thyroglobulin 56 Fcg RIIB mCYT6 FCGR2B 156 Persephin mCYT8 PSPN 57 Cathepsin B CTSB P07858 cathepsin B 157 ICAM-1 ICAM1 P05362 intercellular adhesion molecule 1 257 MCP-3 CCL7 P80098 chemokine (C-C motif) ligand 7 357 Tie-1 TIE1 P35590 tyrosine kinase with immunoglobulin-like and EGF-like domains 1 57 Fetuin A mCYT7 AHSG 157 PF-4 mCYT5 CXCL14 58 Cathepsin L CTSL P07711 cathepsin L 158 ICAM-2 ICAM2 P13598 intercellular adhesion molecule 2 258 MCP-4 CCL13 Q99616 chemokine (C-C motif) ligand 13 358 Tie-2 TEK Q02763 TEK tyrosine kinase, endothelial 58 Flt-3L mCYT6 FLT3LG 158 PlGF-2 mCYT7 Pgf 3. Normal human serum samples were tested with these Sample Name IL-2 IL-4 IL-5 IL-6 IL-8 IL-10 IL-13 GM-CSF IFNg TNFa 59 CCL28 CCL28 Q9NRJ3 chemokine (C-C motif) ligand 28 159 ICAM-3 ICAM3 P32942 intercellular adhesion molecule 3 259 MCSF CSF1 P09603 colony stimulating factor 1 (macrophage) 359 TIM-1 HAVCR1 Q96D42 hepatitis A virus cellular receptor 1 59 Fractalkine mCYT4 CX3CL1 159 Progranulin mCYT7 GRN 60 CD14 CD14 P08571 CD14 molecule 160 IFNab R2 IFNAR2 P48551 interferon (alpha, beta and omega) receptor 2 260 MDC CCL22 O00626 chemokine (C-C motif) ligand 22 360 TIMP-1 TIMP1 P01033 TIMP metallopeptidase inhibitor 1 60 Galectin-1 mCYT6 LGALS1 160 Prolactin mCYT4 PRL 61 CD163 CD163 Q86VB7 CD163 molecule 161 IFNg IFNG P01579 interferon, gamma 261 Mer MERTK Q12866 c-mer proto-oncogene tyrosine kinase 361 TIMP-2 TIMP2 P16035 TIMP metallopeptidase inhibitor 2 61 Galectin-3 mCYT6 LGALS3 161 Pro-MMP-9 mCYT4 Mmp9 Media control 0.1 0.0 0.2 0.1 0.0 0.1 0.1 0.1 0.3 0.1 62 CD200 CD200 P41217 CD200 molecule 162 IGFBP-1 IGFBP1 P08833 insulin-like growth factor binding protein 1 262 Mesothelin MSLN Q13421 mesothelin 362 TIMP-4 TIMP4 Q99727 TIMP metallopeptidase inhibitor 4 62 Galectin-7 mCYT8 LGALS7 162 Prostasin mCYT7 PRSS8 40 Haman inflammation array 63 CD23 FCER2 P06734 Fc fragment of IgE, low affinity II, receptor for (CD23) 163 IGFBP-2 IGFBP2 P18065 insulin-like growth factor binding protein 2, 36kDa 263 MICA MICA Q29983 MHC class I polypeptide-related sequence A 363 Tissue Factor F3 P13726 coagulation factor III (thromboplastin, tissue factor) 63 Gas 1 mCYT6 GAS1 163 P-selectin mCYT4 SELP 64 CD30 TNFRSF8 P28908 tumor necrosis factor receptor superfamily, member 8 164 IGFBP-3 IGFBP3 P17936 insulin-like growth factor binding protein 3 264 MICB MICB Q29980 MHC class I polypeptide-related sequence B 364 TLR2 TLR2 O60603 toll-like receptor 2 64 Gas 6 mCYT6 Gas6 164 RAGE mCYT6 MOK Cell culture, 24hr 0.1 0.9 0.0 0.1 435.0 0.2 0.1 349.7 0.1 0.1 65 CD40 CD40 P25942 CD40 molecule, TNF receptor superfamily member 5 165 IGFBP-4 IGFBP4 P22692 insulin-like growth factor binding protein 4 265 MIF MIF P14174 macrophage migration inhibitory factor (glycosylation-inhibiting factor) 365 TNF RI TNFRSF1A P19438 tumor necrosis factor receptor superfamily, member 1A 65 G-CSF mCYT5 CSF3 165 RANTES mCYT5 CCL5 arrays, where about two thirds of these protein 66 CD40 L CD40LG P29965 CD40 ligand 166 IGFBP-5 IGFBP5 P24593 insulin-like growth factor binding protein 5 266 MIG CXCL9 Q07325 chemokine (C-X-C motif) ligand 9 366 TNF RII TNFRSF1B P20333 tumor necrosis factor receptor superfamily, member 1B 66 GITR mCYT4 TNFRSF18 166 Renin 1 mCYT7 REN 40 Human growth factor array 67 CD97 CD97 P48960 CD97 molecule 167 IGFBP-6 IGFBP6 P24592 insulin-like growth factor binding protein 6 267 MIP-1a CCL3 P10147 chemokine (C-C motif) ligand 3 367 TNFa TNF P01375 tumor necrosis factor 67 GITR L mCYT6 TNFSF18 167 Resistin mCYT4 RETN 68 CEA CEACAMP5 carcinoembryonic antigen-related cell adhesion molecule pseudogene 5 168 IGF-I IGF1 P05019 insulin-like growth factor 1 (somatomedin C) 268 MIP-1b CCL4 P13236 chemokine (C-C motif) ligand 4 368 TNFb LTA P01374 lymphotoxin alpha 68 GM-CSF mCYT5 CSF2 168 SCF mCYT4 KITLG Add drug A, 24hr 0.1 2.4 0.4 0.4 1112.7 0.5 0.0 2399.7 0.1 1.0 69 CEACAM-1 CEACAM1 P13688 carcinoembryonic antigen-related cell adhesion molecule 1 169 IGF-I R IGF1R P08069 insulin-like growth factor 1 receptor 269 MIP-1d CCL15 Q16663 chemokine (C-C motif) ligand 15 369 tPA PLAT P00750 plasminogen activator, tissue 69 gp130 mCYT8 Il6st 169 SDF-1a mCYT4 CXCL12 40 Human chemokine array Mouse array 4 40 70 Chemerin RARRES2 Q99969 retinoic acid receptor responder (tazarotene induced) 2 170 IGF-II Igf2 P01344 insulin-like growth factor 2 (somatomedin A) 270 MIP-3a CCL20 P78556 chemokine (C-C motif) ligand 20 370 TPO THPO P40225 thrombopoietin 70 Granzyme B mCYT8 GZMB 170 sFRP-3 mCYT8 FRZB 71 CHI3L1 CHI3L1 P36222 chitinase 3-like 1 (cartilage glycoprotein-39) 171 IGF-II R IGF2R P11717 insulin-like growth factor 2 receptor 271 MIP-3b CCL19 Q99731 chemokine (C-C motif) ligand 19 371 TRAIL R1 TNFRSF10A O00220 tumor necrosis factor receptor superfamily, member 10a 71 Gremlin mCYT8 GREM1 171 Shh-N mCYT8 SHH Add drug B, 24hr 0.0 1.6 0.1 0.2 576.7 0.2 0.0 493.0 0.1 0.2 72 Ck beta 8-1 CCL23 P55773 chemokine (C-C motif) ligand 23 172 IL-1 R4 IL1RL1 Q01638 interleukin 1 receptor-like 1 272 MMP-1 MMP1 P03956 matrix metallopeptidase 1 (interstitial collagenase) 372 TRAIL R2 TNFRSF10B O14763 tumor necrosis factor receptor superfamily, member 10b 72 H60 mCYT7 H60a 172 SLAM mCYT8 SLAMF1 40 Human receptor array Mouse array 5 40 73 Clusterin CLU P10909 clusterin 173 IL-1 R6 IL1RL2 Q9HB29 interleukin 1 receptor-like 2 273 MMP-10 MMP10 P09238 matrix metallopeptidase 10 (stromelysin 2) 373 TRAIL R3 TNFRSF10C O14798 tumor necrosis factor receptor superfamily, member 10c 73 HAI-1 mCYT6 SPINT1 173 TACI mCYT6 TNFRSF13B concentrations are within quantifiable range. 74 CNTF CNTF P26441 ciliary neurotrophic factor 174 IL-1 RI IL1R1 P14778 interleukin 1 receptor, type I 274 MMP-13 MMP13 P45452 matrix metallopeptidase 13 (collagenase 3) 374 TRAIL-R4 TNFRSF10D Q9UBN6 tumor necrosis factor receptor superfamily, member 10d 74 HGF mCYT4 HGF 174 TARC mCYT5 CCL17 75 Cripto-1 TDGF1 P13385 teratocarcinoma-derived growth factor 1 175 IL-1 RII IL1R2 P27930 interleukin 1 receptor, type II 275 MMP-2 MMP2 P08253 matrix metallopeptidase 2 (gelatinase A, 72kDa type IV collagenase) 375 TRANCE TNFSF11 O14788 tumor necrosis factor (ligand) superfamily, member 11 75 HGF R mCYT6 MET 175 TCA-3 mCYT5 CCL1 Add drug A and B, 24hr 0.1 4.6 0.4 0.8 934.9 1.2 0.2 3283.0 0.0 2.4 76 CRP CRP P02741 C-reactive protein, pentraxin-related 176 IL-10 IL10 P22301 interleukin 10 276 MMP-3 MMP3 P08254 matrix metallopeptidase 3 (stromelysin 1, progelatinase) 376 Transferrin TF P02787 transferrin 76 ICAM-1 mCYT5 ICAM1 176 TCK-1 mCYT8 Ppbp 40 Human array 4 Mouse array 6 40 77 CRTAM CRTAM O95727 cytotoxic and regulatory T cell molecule 177 IL-10 Ra IL10RA Q13651 interleukin 10 receptor, alpha 277 MMP-7 MMP7 P09237 matrix metallopeptidase 7 (matrilysin, uterine) 377 Trappin-2 PI3 P19957 peptidase inhibitor 3, skin-derived 77 IFNg mCYT5 IFNG 177 TECK mCYT8 CCL25 78 CTACK CCL27 Q9Y4X3 chemokine (C-C motif) ligand 27 178 IL-10 Rb IL10RB Q08334 interleukin 10 receptor, beta 278 MMP-8 MMP8 P22894 matrix metallopeptidase 8 (neutrophil collagenase) 378 TREM-1 TREM1 Q9NP99 triggering receptor expressed on myeloid cells 1 78 IFN-γ R1 mCYT8 IFNGR1 178 Testican 3 mCYT7 SPOCK1 Cell culture, 72hr 0.1 0.5 0.0 0.2 453.7 0.8 0.3 563.8 0.1 0.6 79 CTLA4 Ctla4 P16410 cytotoxic T-lymphocyte-associated protein 4 179 IL-11 IL11 P20809 interleukin 11 279 MMP-9 MMP9 P14780 matrix metallopeptidase 9 (gelatinase B, 92kDa type IV collagenase) 379 Troponin I TNNI3 P19429 troponin I type 3 (cardiac) 79 IGFBP-2 mCYT4 IGFBP2 179 TGFb1 mCYT8 TGFB1 40 Human array 5 Mouse array 7 40 80 CXCL14 CXCL14 O95715 chemokine (C-X-C motif) ligand 14 180 IL-12p40 IL12B P29460 interleukin 12B (natural killer cell stimulatory factor 2, p40) 280 MPIF-1 CCL23 P55773 chemokine (C-C motif) ligand 23 380 TSH TSHB P01222 thyroid stimulating hormone, beta 80 IGFBP-3 mCYT4 IGFBP3 180 TIM-1 mCYT7 HAVCR1 81 CXCL16 CXCL16 Q9H2A7 chemokine (C-X-C motif) ligand 16 181 IL-12p70 IL12A P29459 interleukin 12A (natural killer cell stimulatory factor 1, p35) 281 MSP a MST1 P26927 macrophage stimulating 1 (hepatocyte growth factor-like) 381 TSLP TSLP Q969D9 thymic stromal lymphopoietin 81 IGFBP-5 mCYT4 IGFBP5 181 TNF RI mCYT5 TNFRSF1A 82 Cystatin C CST3 P01034 cystatin C 182 IL-13 IL13 P35225 interleukin 13 282 NAP-2 Ppbp P02775 pro-platelet basic protein (chemokine (C-X-C motif) ligand 7) 382 TSP-1 THBS1 P07996 thrombospondin 1 82 IGFBP-6 mCYT4 IGFBP6 182 TNF RII mCYT5 TNFRSF1B Add drug A, 72hr 0.2 34.3 0.3 2.4 915.7 6.7 0.4 3304.8 0.3 20.8 83 DAN PARN O95453 poly(A)-specific ribonuclease 183 IL-13 R1 IL13RA1 P78552 interleukin 13 receptor, alpha 1 283 NCAM-1 NCAM1 P13591 neural cell adhesion molecule 1 383 TSP-2 THBS2 P35442 thrombospondin 2 83 IGF-I mCYT4 IGF1 183 TNFa mCYT5 TNF 40 Human array 6 Mouse array 8 40 84 DcR3 TNFRSF6B O95407 tumor necrosis factor receptor superfamily, member 6b, decoy 184 IL-13 R2 IL13RA2 Q14627 interleukin 13 receptor, alpha 2 284 Neprilysin MME P08473 membrane metallo-endopeptidase 384 TWEAK TNFSF12 O43508 tumor necrosis factor (ligand) superfamily, member 12 84 IL-1 R4 mCYT6 IL1RL1 184 TPO mCYT4 THPO 4. This system provides an invaluable tool for immunology 85 Decorin DCN P07585 decorin 185 IL-15 IL15 P40933 interleukin 15 285 NGF R NGFR P08138 nerve growth factor receptor 385 Ubiquitin+1 RPS27A P62979 ribosomal protein S27a 85 IL-10 mCYT5 IL10 185 TRAIL mCYT7 TNFSF10 Add drug B, 72hr 0.2 1.1 0.1 0.3 454.0 0.7 0.3 721.4 0.1 0.3 86 DKK-1 DKK1 O94907 dickkopf WNT signaling pathway inhibitor 1 186 IL-16 IL16 Q14005 interleukin 16 286 Nidogen-1 NID1 P14543 nidogen 1 386 uPA PRAP1 Q96NZ9 proline-rich acidic protein 1 86 IL-12p40 mCYT5 IL12B 186 TRANCE mCYT8 TNFSF11 87 DKK-3 DKK3 Q9UBP4 dickkopf WNT signaling pathway inhibitor 3 187 IL-17 IL17A Q16552 interleukin 17A 287 Notch-1 NOTCH1 P46531 notch 1 387 uPAR PLAUR Q03405 plasminogen activator, urokinase receptor 87 IL-12p70 mCYT4 IL12A 187 TREM-1 mCYT6 TREM1 40 Human array 7 88 Dkk-4 DKK4 Q9UBT3 dickkopf WNT signaling pathway inhibitor 4 188 IL-17B IL17B Q9UHF5 interleukin 17B 288 NOV NOV P48745 nephroblastoma overexpressed 388 VCAM-1 VCAM1 P19320 vascular cell adhesion molecule 1 88 IL-13 mCYT5 IL13 188 TremL1 mCYT8 TREML1 Add drug A and B, 72hr 0.1 60.5 0.5 4.1 810.6 8.1 0.7 3665.1 1.0 44.1 89 DLL1 DLL1 O00548 delta-like 1 (Drosophila) 189 IL-17B R IL17RB Q9NRM6 interleukin 17 receptor B 289 NrCAM NrCAM Q92823 neuronal cell adhesion molecule 389 VE-Cadherin CDH5 P33151 cadherin 5, type 2 (vascular endothelium) 89 IL-15 mCYT5 IL15 189 TROY mCYT6 TNFRSF19 90 DPPIV DPP4 P27487 dipeptidyl-peptidase 4 190 IL-17C IL17C AF152099 interleukin 17C 290 NRG-b1 NRG1 Q02297 neuregulin 1 390 VEGF VEGFA P15692 vascular endothelial growth factor A 90 IL-17 mCYT5 IL17A 190 Tryptase ε mCYT7 PRSS22 40 Human array 8 91 DR6 TNFRSF21 O75509 tumor necrosis factor receptor superfamily, member 21 191 IL-17E IL25 Q9H293 interleukin 25 291 NSE ENO2 P09104 enolase 2 (gamma, neuronal) 391 VEGF R1 Flt1 P17948 fms-related tyrosine kinase 1 91 IL-17B mCYT8 IL17B 191 TSLP mCYT6 TSLP research and for biomarker discovery. 92 Dtk TYRO3 Q06418 TYRO3 protein tyrosine kinase 192 IL-17F IL17F Q96PD4 interleukin 17F 292 NT-3 NTF3 P20783 neurotrophin 3 392 VEGF R2 KDR P35968 kinase insert domain receptor (a type III receptor tyrosine kinase) 92 IL-17B R mCYT8 IL17RB 192 TWEAK mCYT8 TNFSF12 93 E-Cadherin CDH1 P12830 cadherin 1, type 1, E-cadherin (epithelial) 193 IL-17R IL17RA Q96F46 interleukin 17 receptor A 293 NT-4 NTF4 P34130 neurotrophin 4 393 VEGF R3 FLT4 P35916 fms-related tyrosine kinase 4 93 IL-17E mCYT4 IL25 193 TWEAK R mCYT6 TNFRSF12A 94 EDA-A2 EDA Q92838 ectodysplasin A 194 IL-18 BPa IL18BP O95998 interleukin 18 binding protein 294 OPG TNFRSF11B O00300 tumor necrosis factor receptor superfamily, member 11b 394 VEGF-C VEGFC P49767 vascular endothelial growth factor C 94 IL-17F mCYT4 IL17F 194 VCAM-1 mCYT4 VCAM1 40 Human array 9 95 EGF EGF P01133 epidermal growth factor 195 IL-18 Rb IL18RAP O95256 interleukin 18 receptor accessory protein 295 OPN SPP1 P10451 secreted phosphoprotein 1 395 VEGF-D FIGF O43915 c-fos induced growth factor (vascular endothelial growth factor D) 95 IL-1a mCYT5 IL1A 195 VEGF mCYT4 VEGFA 96 EGF R EGFR P00533 epidermal growth factor receptor 196 IL-18 IL18 Q14116 interleukin 18 (interferon-gamma-inducing factor) 296 OSM OSM P13725 oncostatin M 396 Vitronectin VTN P04004 vitronectin 96 IL-1b mCYT5 IL1B 196 VEGF R1 mCYT6 Flt1 Data extraction Individual cytokine concentrations 97 EG-VEGF PROK1 P58294 prokineticin 1 197 IL-1a IL1A P01583 interleukin 1, alpha 297 Osteoactivin GPNMB Q14956 glycoprotein (transmembrane) nmb 397 vWF VWF P04275 von Willebrand factor 97 IL-1ra mCYT4 IL1RN 197 VEGF R3 mCYT6 FLT4 98 EMMPRIN BSG P35613 basigin 198 IL-1b IL1B P01584 interleukin 1, beta 298 PAI-I SERPINE1 P05121 serpin peptidase inhibitor, clade E, member 1 398 WIF-1 WIF1 Q9Y5W5 WNT inhibitory factor 1 98 IL-2 mCYT5 IL2 198 VEGF-B mCYT8 VEGFB 99 ENA-78 CXCL5 P42830 chemokine (C-X-C motif) ligand 5 199 IL-1F10 IL1F10 Q8WWZ1 interleukin 1 family, member 10 (theta) 299 PAPP-A PAPPA Q13219 pregnancy-associated plasma protein A, pappalysin 1 399 WISP-1 WISP1 O95388 WNT1 inducible signaling pathway protein 1 99 IL-2 Ra mCYT4 IL2RA 199 VEGF-D mCYT4 FIGF 100 Endoglin ENG P17813 endoglin 200 IL-1F5 IL36RN Q9UBH0 interleukin 36 receptor antagonist 300 PARC CCL18 P55774 chemokine (C-C motif) ligand 18 (pulmonary and activation-regulated) 400 XEDAR EDA2R Q9HAV5 ectodysplasin A2 receptor 100 IL-20 mCYT4 IL20 200 VEGF-R2 mCYT8 KDR Human Multiplex ELISA Arrays (400) Mouse Multiplex ELISA Arrays (200)