KRIBIOLISA™ Rat Anti-Ranibizumab (Lucentis) ELISA

Krishgen- Biopharma-PK-ADA; Biosimilars / Biotherapeutics; Immunogenecity Assays

Enzyme Immunoassay for the quantitative determination of Anti-Ranibizumab in serum and plasma

Introduction:
Ranibizumab (Lucentis) is a recombinant humanized IgG1 monoclonal antibody fragment that binds to and
inhibits vascular endothelial growth factor A (VEGF-A). VEGF is a biochemical signal protein that promotes
angiogenesis throughout the body and in the eye. Through binding to VEGF-A, ranibizumab interrupts the
interaction of VEGF with its receptors, and thus prevents the subsequent growth of new blood vessels.
Anti-Drug Antibodies (ADA) may induce unwanted side effects in biopharmaceuticals. Hence, ADA has been
subjected to increase in scrutiny by the regulatory authorities using immunogenicity safety studies. ADA has
been observed in pre-clinical and clinical studies, resulting in significant changes in toxicology,
pharmacokinetics and efficacy. These effects result from the generation of drug-induced (neutralizing)
autoantibodies against Ranibizumab and can be responsible for allergic reaction, or even anaphylactic shock.
This ELISA kit detects antibodies for Anti-Ranibizumab and may be used for monitoring immunogenicity.

Intended Use:
The KRIBIOLISA™ Rat Anti-Ranibizumab ELISA is used as an analytical tool for quantitative determination of
Anti-Ranibizumab in rat serum.

Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Ranibizumab is pre-coated
onto microwells. Samples and standards are pipetted into microwells and antibodies to Ranibizumab present in
the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated Ranibizumab
is pipetted and incubated. After washing microwells in order to remove any nonspecific binding, the ready to use
substrate solution (TMB) is added to microwells and color develops proportionally to the amount of AntiRanibizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is
measured at 450 nm.