Abstract |
The fully humanized Lewis-Y carbohydrate specific monoclonal antibody (mAb) IGN311 is currently tested in a passive immunotherapy approach in a clinical phase I trail and therefore regulatory requirements demand qualified assays for product analysis. To demonstrate the functionality of its Fc-region, the capacity of IGN311 to mediate complement dependent cytotoxicity (CDC) against human breast cancer cells was evaluated. The "classical" radioactive method using chromium-51 and a FACS-based assay were established and qualified according to ICH guidelines. Parameters evaluated were specificity, response function, bias, repeatability (intra-day precision), intermediate precision (operator-time different), and linearity (assay range). In the course of a fully nested design, a four-parameter logistic equation was identified as appropriate calibration model for both methods. For the radioactive assay, the bias ranged from -6.1% to -3.6%. The intermediate precision for future means of duplicate measurements revealed values from 12.5% to 15.9% and the total error (beta-expectation tolerance interval) of the method was found to be <40%. For the FACS-based assay, the bias ranged from -8.3% to 0.6% and the intermediate precision for future means of duplicate measurements revealed values from 4.2% to 8.0%. The total error of the method was found to be <25%. The presented data demonstrate that the FACS-based CDC is more accurate than the radioactive assay. Also, the elimination of radioactivity and the 'real-time' counting of apoptotic cells further justifies the implementation of this method which was subsequently applied for testing the influence of storage at 4 degrees C and 25 degrees C ('stability testing') on the potency of IGN311 drug product. The obtained results demonstrate that the qualified functional assay represents a stability indicating test method.
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Authors | A Nechansky, O H J Szolar, P Siegl, I Zinoecker, N Halanek, S Wiederkum, R Kircheis |
Journal | Journal of pharmaceutical and biomedical analysis
(J Pharm Biomed Anal)
Vol. 49
Issue 4
Pg. 1014-20
(May 01 2009)
ISSN: 1873-264X [Electronic] England |
PMID | 19250790
(Publication Type: Comparative Study, Journal Article)
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Chemical References |
- Antibodies
- Antibodies, Monoclonal
- Coloring Agents
- IGN 311
- Lewis Blood Group Antigens
- Lewis Y antigen
- Complement System Proteins
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Topics |
- Algorithms
- Antibodies
(toxicity)
- Antibodies, Monoclonal
(immunology)
- Biological Assay
- Cell Line, Tumor
- Cell Survival
- Coloring Agents
- Complement System Proteins
(physiology)
- Data Interpretation, Statistical
- Female
- Flow Cytometry
- Humans
- Lewis Blood Group Antigens
(immunology)
- Linear Models
- Reproducibility of Results
- Risk Assessment
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