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Antibody Screening by Microarray Technology—Direct Identification of Selective High-Affinity Clones

Federal Institute for Materials Research and Testing (BAM), Division 1.5 Protein Analysis, Richard-Willstätter-Strasse 11, 12489 Berlin, Germany
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Antibodies 2020, 9(1), 1; https://doi.org/10.3390/antib9010001
Received: 1 November 2019 / Revised: 9 December 2019 / Accepted: 11 December 2019 / Published: 2 January 2020
The primary screening of hybridoma cells is a time-critical and laborious step during the development of monoclonal antibodies. Often, critical errors occur in this phase, which supports the notion that the generation of monoclonal antibodies with hybridoma technology is difficult to control and hence, a risky venture. We think that it is crucial to improve the screening process to eliminate most of the critical deficits of the conventional approach. With this new microarray-based procedure, several advances could be achieved: Selectivity for excellent binders, high-throughput, reproducible signals, avoidance of misleading avidity (multivalency) effects, and performance of simultaneous competition experiments. The latter can also be used to select clones of desired cross-reactivity properties. In this paper, a model system with two excellent clones against carbamazepine, two weak clones, and blank supernatant containing fetal bovine serum was designed to examine the effectiveness of the new system. The excellent clones could be detected largely independent of the immunoglobulin G (IgG) concentration, which is usually unknown during the clone screening since the determination and subsequent adjustment of the antibody concentration are not feasible in most cases. Furthermore, in this approach, the enrichment, isolation, and purification of IgG for characterization is not necessary. Raw cell culture supernatant can be used directly, even when fetal calf serum (FCS) or other complex media is used. In addition, an improved method for the oriented antibody-immobilization on epoxy-silanized slides is presented. Based on the results of this model system with simulated hybridoma supernatants, we conclude that this approach should be preferable to most other protocols leading to many false positives, causing expensive and lengthy elimination steps to weed out the poor clones. View Full-Text
Keywords: monoclonal antibodies; mabs; fusion; false positives; hapten immunoassays; competitive immunoassays; ELISA; antibody validation; antibody quality; microarray; hybridoma technology; linker recognition; high-throughput screening; HTS; heterology concept monoclonal antibodies; mabs; fusion; false positives; hapten immunoassays; competitive immunoassays; ELISA; antibody validation; antibody quality; microarray; hybridoma technology; linker recognition; high-throughput screening; HTS; heterology concept
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MDPI and ACS Style

Paul, M.; Weller, M.G. Antibody Screening by Microarray Technology—Direct Identification of Selective High-Affinity Clones. Antibodies 2020, 9, 1.

AMA Style

Paul M, Weller MG. Antibody Screening by Microarray Technology—Direct Identification of Selective High-Affinity Clones. Antibodies. 2020; 9(1):1.

Chicago/Turabian Style

Paul, Martin; Weller, Michael G. 2020. "Antibody Screening by Microarray Technology—Direct Identification of Selective High-Affinity Clones" Antibodies 9, no. 1: 1.

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