BODITECH MED INC. I-CHROMA AFP-25

BODITECH MED INC. i-CHROMA AFP-25

BodiTech Med’s express and implied warranties (including implied warranties of merchantability and fitness) are conditional upon observance of BodiTech Med’s published directions with respect to the use of BodiTech Med’s products.

For Technical Assistance call Boditech Med’s Technical Services at Tel: +82 (33) 243-1400 E-mail: [email protected]

BodiTech Med Incorporated

G-Tech Village, 1144-2 Geoduri Dongnaemyeon, Chuncheon, Gangwondo 200-883 Republic of Korea Tel: +82 (33) 243-1400

Fax: +82 (33) 243-9373 www.boditech.co.kr

EU Representative: Jai Jun Choung, Ph.D. EU Biotech Development Ltd. 81 Oxford Street, LONDON, W1D 2EU United Kingdom Tel: +44 207 903 5441 Fax: +44 207 903 5333 E-Mail: [email protected]

i-CHROMATM is registered trademarks of BodiTech Med Incorporated.

Revision No: 05 Date of last revision: May 13, 2010.

BodiTech Med Inc.

Rev. 05_100513_M BODITECH MED INC. i-CHROMA AFP-25

The i-CHROMATM AFP Test Kit consists of Test Device and TM Detector Buffer. Test Device is individually sealed with a desiccant i-CHROMA AFP in aluminum pouch, and Detector Buffer is packed and delivered separately from Test Device in a styrofoam box filled with ice pack. ImmunoAssay for Quantitative Measurement of Alpha Feto Protein(AFP) in Human Whole blood with i- • Test Device contains a test strip in which murine monoclonal CHROMATM Reader System. antibody against human AFP and rabbit IgG have been immobilized on the test and on the control line of strip, respectively. • Detector Buffer contains fluorescence-labeled anti-AFP (Mouse INTENDED USE monoclonal), fluorescence-labeled anti-rabbit IgG, BSA as a The i-CHROMATM AFP Test along with i-CHROMATM Reader is stabilizer, and Sodium Azide as a preservative in PBS. a fluorescence immunoassay that measures AFP in human whole blood. WARNINGS AND PRECAUTIONS • IVD For In Vitro Diagnostic Use. INTRODUCTION • Carefully follow the instructions and procedures described in this Alpha-fetoprotein (AFP) is a 1-globulin family of human insert. REF Catalog No. i-CHROMATM AFP-25 plasma proteins and a glycoprotein with a molecular weight approximately 70 kDa. AFP is produced primarily in the • Don’t use Test Device if its lot # does not match with ID Chip # liver of developing fetus. It can be found in maternal blood that is inserted onto the instrument. and in amniotic fluid since it is secreted into fetal serum. • The i-CHROMATM AFP Test Kit is only operational in the i- The concentration of AFP in healthy adult is below 15 ng/ml CHROMATM Reader. And tests should be applied by but it shows a great increase in several malignant diseases, professionally trained staff working in certified laboratories at mostly primary hepatocelluar carcinoma and non- some remove from the patient and clinic at which the sample(s) seminomatous testicular cancer. Some 70-90% of patients is taken by qualified medical personnel. with primary hepatocelluar carcinoma and • LOT Neither inter-change materials from different product lots nonseminomatous testicular cancer have been observed to nor use beyond the expiration date. The use of medical have high levels of serum AFP. High levels of serum AFP device beyond expiration date may affect on test result. also have been found in a limited number of patients TM diagnosed with various diseases such as gastrointestinal tract • The i-CHROMA AFP Test Device should remain in its cancer, viral hepatitis, chronic active hepatitis, alcoholic original sealed pouch until ready to use. Do not use the Test Device if the pouch is damaged or the seal is broken. cirrhosis, and adenocarcinomas of lung, pancreas, and gall Discard after single use. bladder. Since AFP is well known to be an important prognostic indicator of non-seminomatous testicular cancer, • AFP Test Device and Reader should be used away from vibration its most definitive role is on monitoring post-treatment and magnetic field. During normal usage, i-CHROMATM AFP clinical status and the post therapeutic evaluation of patients. Test may introduce minute vibration, which should be regarded normal. • Use separate clean pipette tips and sample vials for different PRINCIPLE specimens. The pipette tips and sample vials should be used for one specimen only. Discard after single use. The i-CHROMATM AFP Test is based on fluorescence immunoassay technology. The i-CHROMATM AFP Test uses a • Blood specimens, used test devices, pipette tips and sample vials sandwich immunodetection method, such that by mixing detector are potentially infectious. Proper laboratory safety buffer with blood specimen in test vial, the fluorescence-labeled techniques, handling and disposal methods should be detector anti-AFP antibody in buffer binds to AFP antigen in blood followed in accordance with standard procedures and specimen. As the sample mixture is loaded onto the sample well of relevant regulations observed by microbiological hazard the test device and migrates the nitrocellulose matrix of test strip by materials. capillary action, the complexes of detector antibody and AFP are captured to anti-AFP sandwich pair antibody that has been • The test will be applied on a routine basis and not in emergency immobilized on test strip. Thus the more AFP antigen is in blood situations. specimen, the more complexes are accumulated on test strip. Signal • Do not smoke, eat, or drink in areas in which specimens or kit intensity of fluorescence of detector antibody reflects amount of reagents are handled. AFP captured and is microprocessed from i-CHROMATM Reader to show AFP concentration in blood specimen. The default result unit TM of i-CHROMA AFP Test is displayed as an ng/L from i- STROAGE AND STABILITY CHROMATM Reader. The working range and the detection limit of TM i-CHROMA AFP Test system are 5 - 350 ng/mL and 5ng/mL • Store the detector buffer in a refrigerator at 2° - 8°C. The respectively. Detector Buffer is stable up to 20 months if stored in a * Reference Value : 20 ng/mL refrigerator. COMPOSITION OF REAGENTS

• Once removed from refrigerator, allow the Detector Buffer for 30 minutes to return to room temperature before testing. • Store i-CHROMATM AFP Test Device at 4°-30°C in its sealed pouch. The i-CHROMATM AFP Test Device is stable for 20 months (while in the sealed pouch) if stored at 4°-30°C. • If stored in a refrigerator, allow a minimum of 30 minutes for the Test Device to reach room temperature while it is in the sealed pouch. • Do not remove the device from the pouch until ready to use. The Test Device should be used immediately once opened. • The storage and shipping of Test Kit should be complied as indicated in manual. However, it is remotely possible that only part of Test Kit is affected by stability problems.

SAMPLE COLLECTION AND PREPARATION The test can be performed with either serum or plasma or whole blood. • For serum sample, collect the blood in a tube without anticoagulant and allow to be clotted. Remove the serum from the clot as soon as possible to avoid hemolysis. For plasma sample, collect the blood in a tube treated with EDTA. Anticoagulants other than EDTA for plasma specimen have not been evaluated. If testing cannot be conducted within an hour after preparation of specimen, the serum/plasma should be stored at -20o C until tested. In case of use whole blood, apply it immediately after specimen was taken. • The specimen must be at room temperature and be homogeneous before testing. Frozen specimens must be completely thawed, thoroughly mixed, and brought to room temperature prior to testing. If specimens are to be shipped, they should be packed in compliance with regulations. • It is recommended to avoid using severely hemolyzed specimens whenever possible. If a specimen appears to be severely hemolyzed, another specimen should be obtained and tested.

MATERIALS PROVIDED BodiTech Med Incorporated i-CHROMATM AFP Test REF Catalog No. i-CHROMATM AFP -25

Kit contains: Test Devices 25T/box Detector Buffer 1vial (2ml/vial) ID Chip 1/box Insert 1 ea

MATERIALS REQUIRED BUT NOT PROVIDED i-CHROMATM Reader REF Catalog No. FR-203 Thermal Printer Transfer pipette (10, 20, 75µL size)

about obtaining the controls, contact BodiTech Med PROCEDURE Incorporate’s Technical Services for assistance. • Image of the test kit Procedure Control  Each i-CHROMATM AFP Test Device contains internal control that satisfies routine quality control requirements. This internal window sample well control is performed each time a patient sample is tested. This control indicates that the test device was inserted and read TM 1. Set a Test Device on a dust-free clean place. properly by i-CHROMA Reader. An invalid result from the internal control causes an error message on i-CHROMATM 2. Check/insert ID Chip onto the instrument. Make sure Reader indicating that the test should be repeated. that the Test Device lot # matches with ID Chip #. 3. Take out one tube of Detection Buffer from refrigerator and leave it at room temperature. LIMITATIONS OF THE PROCEDURE TM 4. Draw 30 µL of whole blood (15 µL of serum, plasma or • The results of i-CHROMA AFP Test should be evaluated with Control) with a transfer pipette and add it to the tube all clinical and laboratory data available. If AFP Test results do containing Detector Buffer. not agree with the clinical evaluation, additional tests should be performed. 5. Mix well the specimen with Detector Buffer by tapping or inverting the tube. • The false positive results include cross-reactions with some components of serum from individual to antibodies; and non- 6. Take 75 µL of sample mixture and load it onto the specific adhesion of some components in human blood that have well of disposable Test Device. similar epitopes to capture and detector antibodies. In the case of 7. Leave the Test Device at room temperature for 15 min before false negative results, the most common factors are: non- inserting the device into the holder. responsiveness of antigen to the antibodies by that certain unknown components are masking its epitope, such that antigen 8. To start scanning, insert test device onto the holder of i- cannot be seen by the antibodies; instability of AFP antigen, CHROMATM Reader and press “SELECT” button.. Make resulting in degradation with time and, or temperature, such that sure direction of Test Device and push the device back they become no longer recognizable by antibodies; and degraded all the way. The instrument will automatically start to scan other test components. The effectiveness of the test is highly the Test Device immediately. dependent on storage of kits and sample specimens at optimal conditions. 9. Read the results on the display screen of i- TM CHROMA Reader. • Plasma using anticoagulants (e.g. heparin or citrate) other than EDTA has not been evaluated in i-CHROMATM AFP Test and  Refer to i-CHROMATM Reader Operation Manual for the thus should not be used. complete instructions on use of the Test. • Other factors may interfere with i-CHROMA TM AFP Test and may cause erroneous results. These include technical or RESULT procedural errors, as well as additional substances in blood specimens. The i-CHROMATM Reader calculates AFP test results automatically and displays AFP concentration on the screen as form of mg/L. For further information, refer to the Operation PERFORMANCE CHARACTERISTICS Manual for the i-CHROMATM Reader. 1. Analytical Sensitivity: Analytical sensitivity means the lowest concentration of AFP that the test system can detect with Quality Control CV<10%. Analytical sensitivity of i-CHROMATM AFP Test was determined by testing 10 times with three lots of reagents. Quality Control Analytical sensitivity of i-CHROMATM AFP Test system was 5 ng/mL.  A quality control test using commercially available controls should be performed as a part of good testing practice, to 2. Specificity: Other bio-molecules, such as Hb, CEA, PSA, confirm the expected QC results, to confirm the validity of the ALT, Troponin I, CK-MB, Albumin, and serum amyloid P assay, and to assure the accuracy of patient results. If you want component were added to test specimen with much higher level to perform QC of Test Kit, we recommend using Abbott Axsym than their physiological level in normal blood. control. 3. Imprecision: For the intra-assay imprecision, 20 replicates were  A quality control test should be performed at regular intervals, tested at each control sample. For the inter-assay imprecision, and before using a new kit with patient specimens, controls tests were conducted on 10 sequential days, with 10 runs per day should be tested to confirm the test procedure, and to verify the and with 10 replicates at each AFP concentration. tests produce the expected QC results. QC specimens should also be run whenever there is any question concerning the validity of Imprecision of i-CHROMATM AFP Test Kit results obtained. Upon confirmation of the expected results, the test device is ready to use with patient specimens. Control standards are not provided with this test kit. For information Intra-assay Inter-assay

AFP Mean S.D CV% Mean S.D CV% (ng/mL)

20 21 1.2 5.7 22 1.3 5.9

50 53 3.5 6.6 52 3.8 7.3

200 202 6.2 6.2 205 11.1 5.4

4. Linearity: The high concentration was diluted with the low concentration to the following final percentages; 100%, 75%, 50%, 25%, 10%, 5% and 0%. Sample was assayed in triplicate in one analytical run at each AFP level. The coefficient of linear regression was R=0.997. Linearity of i-CHROMATM AFP Test Kit was 5~350ng/mL. 5. Comparability: The AFP concentrations of 30 clinical specimens were quantified independently with i-CHROMATM AFP Test Kit and Axsym (Abbott) automatic analyzer. While the whole blood was used for i-CHROMATM AFP Test Kit, the serums were used for Abott Axsym. The test results were compared and their compatibilities were investigated with linear regression and correlation of coefficient (R). i-CHROMATM AFP Test was comparable well to other method (R=0.989).

250 Y=0.90826x-4.43243 R=0.98907 200 SD=9.90543 ) l m /

g 150 n ( P F A

100 A M O

R 50 H C - i 0

0 50 100 150 200 250 Abbott Axsym AFP(ng/ml)

REFERENCES 1. Tartarinov, Y.S. Detection of embryospecific alpha-globulin in the blood sera of patients with primary liver tumor. Vopr. Med. Khim. 10:90-91 (1964). 2. Mcintire, K.R., Waidmann, T.A., Moertel, C.G. and Go, V.L.W. Serum alpha-fetoprotein in patients with neoplasms of the gastrointestinal tract. Cancer Res. 35:991-996 (1975). 3. Javadpouf, N., Mcintire, K.R. and Waidmann, T.A. Human chorionic gonadotropin (HCG) and alpha-fetoprotein (AFP) in sera and tumor cells of patients with testicular seminoma. Cancer 42:2768-2772. (1978). 4. Chen, D.S. and Sung, J.L. Relationship of Hepatitis B Surface Antigen to serum alpha-fetoprotein in nonmalignant diseases of the lever. Cancer 44:984-992 (1979). 5. Rhoslati,E. and Seppala, M. studies of carcinofetal proteins: Physical and Chemical Properties of Human alpha-fetoprotein. Int. J. Cancer 7:218-225 (1971). 6. Abelev,G.I. Alpha-fetoprotein in oncogenesis and its association with malignant tumors. Adv. Cancer Res. 7:295-358 (1971). 7. Wespic, H.C. Alpha-fetoprotein: its quantification and relationship to neoplastic disease, ppp 115-129 In Alpha-fetoprotein, Laboratory Procedures and Clinical Applications, Kirkpatirck, A. and Nakamuram R (eds.), Masson Publishing, New York (1981) After Radical Prostatectomy. J. Urol. 142:1082-90 (1989).

Rev. 05_100513_M