IMMUNOLOGY Exploring Signaling LUNARIS™ Chemokine Kits

Human 10-Plex: CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL20, CXCL1, CXCL8 (IL-8), CXCL10 (IP-10), CXCL12 (SDF1A)

Mouse 11-Plex: CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CCL22, CXCL1, CXCL10 (IP-10), CX3CL1 (Fractalkine)

Chemokines

Chemokine receptor

Plasma membrane its | 2020.01 | 09075 | www.vierviertel.com | 09075 | 2020.01 its Cytoplasm

K Chemokine AYOXXA Intracellular signaling

Chemotaxis

www.ayoxxa.com LUNARIS™ Human & Mouse Chemokine Kits Validated, scalable, robust

Validated for quantitative analysis of up to 12 different in one sample

Profile homeostatic and proinflammatory chemokine signatures in sample volumes as low as 3 µL

Scalable and standardized assay architecture with robust chemistry guarantees reproducibility

Harmonized panels for human and murine samples allow rapid translation of results from lab to clinic

LUNARIS™ Human 10-Plex Chemokine Kit Quantifies CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL20, CXCL1, CXCL8 (IL-8), CXCL10 (IP-10), CXCL12 (SDF1A) in serum and cell culture supernatant.

LUNARIS™ Mouse 11-Plex Chemokine Kit Quantifies CCL2 (MCP-1), CCL3, CCL4, CCL5 (RANTES), CCL11 (Eotaxin), CCL19, CCL20, CCL22, CXCL1, LUNARIS™ Human 10-Plex CXCL10 (IP-10), CX3CL1 (Fractalkine) Chemokine Kit in serum and cell culture supernatant. For Research Use Only. Not for use in diagnostic procedures.

Chemokine Signaling

Chemokines constitute the largest family of and mediate versatile and powerful signaling in cell migration, Analytes contained in LUNARIS™ Chemokine Kits homeostasis, inflammation, , embryogenesis, grouped by cellular function cancer metastasis and several other medically relevant functions.

The four classes of chemokines initiate complex signaling cascades by binding to specific 7-transmembrane G - Lymphocyte Mononuclear Granulocyte coupled receptors. traffic cell traffic traffic

Building fine-tuned concentration gradients, chemokines exert precise control of diverse regulatory pathways and cause pleiotropic effects that extend far beyond their originally described role as leucocyte chemoattractants. A plethora of literature bears witness to the known importance of these small cytokines in a vast number of biological functions, while a growing number of publications hints at a yet undescribed complex interplay between target cells and their respective Th1, Th2, Th17 cells , Eosinophils, chemokines. natural killer cells neutrophils

Chemokine repertoires of some signaling axes are extensively conserved between mice and humans, making murine models the system of choice in more than just immunology contexts. CCL20, CCL22, CCL2, CCL3, CCL4, CCL11, CXCL1, CX3CL1, CXCL10 CCL5, CCL19, CX3CL1 CXCL8 (IL-8) Relevant research areas:

Inflammation and infection biology Note: CXCL12 is mainly involved in homing Allergy and autoimmune disease (e.g., retention of granulocytes, B cells and precursors). Hematopoiesis and angiogenesis

www.ayoxxa.com 2 Excellent Data Quality Validation of LUNARIS™ Human 10-Plex Chemokine Kit*

Table 1: Performance Analyte LoD LLOQ ULOQ DR Figure 1: Performance CCL2 Median assay parameters Bar diagram depicting the CCL2 0.2 0.3 200 2.3 CCL3

from 20 experiments. CCL3 4.6 12.0 300 2.5 median assay parameters CCL4

L0D = Limit of detection, CCL4 2.9 4.8 600 2.8 shown in table 1. The orange CCL5 LLOQ = Lower limit of CCL5 1.4 8.0 1000 3.0 bars represent the dynamic CCL11 quantification, CCL11 2.3 7.2 900 3.0 range of the assay from LoD CCL20 Analytes ULOQ = Upper limit CCL20 5.1 24.0 600 2.8 (left border) to the ULOQ CXCl1 CXCL1 9.2 40.0 1000 3.0 Analytes CXCL8 of quantification, (right border). The LLOQ is CXCL10 CXCL8 0.3 0.5 300 2.5 DR = Logarithmic detection depicted as a white line. CXCL12 CXCL10 4.9 8.0 3000 3.5 range. All values reported Measurements from 20 1 1 0 CXCL12 24.2 120.0 3000 3.5 10 0. 10 1000 0000 in pg/mL. validation experiments. 1 concentration [pg/mL] Cytokine concentration [pg/mL ]

Figure 2: Accuracy Figure 3: Precision 200 40 Median recovery rate Between-run precision (BRP) from 10 experiments of QC samples spiked into cell 150 30 quantifying markers in %) culture supernatants. Each data y( quality control samples point represents the median %)

100 P( 20 ver

spiked at two different between-run precision of (%) BRP BR eco Recovery (%) concentrations into cell R 50 two different QC sample 10 culture supernatants. concentrations measured in The targeted recovery 0 10 experiments. The targeted 0 3 4 5 0 2 2 3 4 5 1 0 2 range (70–130%) is 2 11 20 L1 L8 precision (<20%) is denoted 11 20 L L8 CL CL CL C C L1 L1 CL CL CL CL C L1 L1 CL C C CL CL X X C C C C C C CL CL XC X C C C C C C C X X C C C C X X denoted by the dotted C C C by the dotted lines/grey C C lines/grey shaded area. shaded area.

*Validation data of the corresponding mouse kit are available upon request.

Innovative technology for Multiple options to access translational proteomics the advantages of LUNARIS™

Ease of readout and handling Complete testing and readout services Fully integrated system Send in samples for testing and receive a complete analysis report Flexible scalability from low to high throughput Perform assays in your lab, then send in Readout of 384 samples in less than one hour completed assay plates for readout

AYOXXA’s proprietary LUNARIS™ platform for multiplex protein Innovative panel development analysis is a fully integrated and scalable system. A dedicated Custom panel configuration reader automates image-based analysis of immunoassay beads in a planar array, so every bead is read and every bead Custom multiplex assay development counts.

LUNARIS™ is optimized for sample volumes as low as 3 µL – one-tenth the volume required for similar technologies – yet allows full multiplex testing without compromising data quality or precision from precious samples.

With advantages of superior data quality, workflow flexibility and conservation of precious samples, LUNARIS™ enables reliable quantification of biomarkers from model to man – from lab to clinic – from data to insight.

www.ayoxxa.com 3

Ordering information * Chemokine Kit Human 10-Plex LUNARIS™ Kits Chemokine Kit Mouse 11-Plex Measured induplicate www.ayoxxa.com [email protected] +49 Call theexperts: signaling chemokine Exploring technology for translational proteomics innovative multiplex protein AYOXXA’s www.ayoxxa.com ( 0 ) 221-222 529-0 4 ×96 1 ×96 BioChips of No. 4 ×96 1 ×96 LHCK-20100F LHCK-20100S Cat. No. Cat. LMCK-20110F LMCK-20110S samples of No. 184 40 184 40

* E-Mail: [email protected] E-Mail: 529-0 (0) 222 +49 Phone: 221 50829 1 Nattermannallee Cologne BioCampus GmbH Biosystems AYOXXA lab to clinic of biomarkers model from to man –from quantification reliable enables LUNARIS™ terms of quality, and efficiency, flexibility clinicalresearch. advantages With its in to basic from knowledge translating protein analysis platform is optimized for multiplex LUNARIS™ Our applications. panelsassay for translational research platform, our mission is to develop robust Building technology innovative an upon research. vision of enabling success in translational Biosystems AYOXXA HEADQUARTERS GERMANY Köln, Germany – from data– from to insight. is dedicated to the E-Mail: [email protected] E-Mail: USA 20852 MD Rockville, c/o Platinum CFO c/o Platinum Inc. Biosystems AYOXXA 2600 USA Tower Oaks Blvd., Suite 220 Suite Blvd., Oaks Tower

4

AYOXXA Chemokine Kits | 2020.01 | 09075 | www.vierviertel.com