Abstract

Monitoring host cell protein (HCP) and protein A impurities is important to ensure successful development of recombinant antibody drugs. Here, we report the full automation and validation of an ELISA platform on a robotic system that allows the detection of Chinese hamster ovary (CHO) HCPs and residual protein A of in-process control samples and final drug substance. The ELISA setup is designed to serve three main goals: high sample throughput, high quality of results, and sample handling flexibility. The processing of analysis requests, determination of optimal sample dilutions, and calculation of impurity content is performed automatically by a spreadsheet. Up to 48 samples in three unspiked and spiked dilutions each are processed within 24 h. The dilution of each sample is individually prepared based on the drug concentration and the expected impurity content. Adaptable dilution protocols allow the analysis of sample dilutions ranging from 1:2 to 1:2×10(7). The validity of results is assessed by automatic testing for dilutional linearity and spike recovery for each sample. This automated impurity ELISA facilitates multi-project process development, is easily adaptable to other impurity ELISA formats, and increases analytical capacity by combining flexible sample handling with high data quality.

Links

Publisher Full Text

Authors

Rey G, Wendeler MW

Institution

Novartis Pharma AG, TRD Biologics - Analytical R&D, CH-4002 Basel, Switzerland.

Source

Journal of pharmaceutical and biomedical analysis 70: 2012 Nov pg 580-6

MeSH

Animals
Antibodies
Automation, Laboratory
CHO Cells
Calibration
Cricetinae
Cricetulus
Drug Contamination
Enzyme-Linked Immunosorbent Assay
Limit of Detection
Linear Models
Recombinant Proteins
Reference Standards
Reproducibility of Results
Robotics
Staphylococcal Protein A
Transfection

Pub Type(s)

Journal Article
Validation Studies

Language

eng

PubMed ID

22727805