Antigen generation and display in therapeutic antibody drug discovery – a neglected but critical player

Abstract

Disease intervention by targeting a critical pathway molecule through a blocking antibody or interference by therapeutic proteins is currently en vogue. Generation of blocking antibodies or therapeutic proteins inevitably requires the production of recombinant proteins or cell-based immunogens. Thus, one could call the antigen molecule the neglected player in antibody drug discovery. The variety of methods available for making recombinant proteins or recombinant cell lines that present the target on the cell surface is extensive. These need to be addressed in conjunction with biochemical and biophysical quality criteria and the experimental application intended. Fundamentally, successful production and isolation of monoclonal antibodies requires optimized antigen preparation and presentation to the immune host. This review summarizes the most important aspects of antigen generation and display, enabling logical decision making to give rise to potent high-affinity antibodies.

ATR-FTIR spectroscopy and spectroscopic imaging for the analysis of biopharmaceuticals

“…7). Upstream techniques in processing of biopharmaceuticals experience high levels of monitoring and feedback when compared to downstream techniques [97][98][99]. For example, inline applications of ATR-FTIR spectroscopy include upstream process monitoring of glucose and ethanol in different cell lines, observing changes in α-glucose spectra with time, and the possibility of simplified calibration techniques using a spectral library [100,101].…”

Screening Strategies for the Discovery of Ion Channel Monoclonal Antibodies

“…Ion channels are normally expressed in low abundance in vivo and the heterologous expression of the proteins needed to achieve high expression levels requires considerable experimentation to optimize the host expression system, particularly as heterologous overexpression can be toxic (Vu, Bautos, Hong, & Gelli, 2009). Host cells for heterologous expression include bacteria (e.g., E. coli), insect cells (e.g., Sf9 cells), yeast (e.g., Pichia pastoris), the ciliated protozoan Tetrahymena thermophila, and mammalian cells (Assenberg, Wan, Geisse, & Mayr, 2013;Bisharyan et al, 2014;Ebersbach & Geisse, 2012;Kost & Kemp, 2016). Ion channels with large extracellular domains (ECD) may prove amenable to expression of the ECDs for use as antigens (Nury et al, 2010).…”

Selecting an Optimal Antibody for Antibody- Drug Conjugate Therapy

“…The choice of host likely depends on the nature of target molecule. Most importantly, the yield should be a homogenous, properly folded antigen such that the critical epitopes are accessible (Ebersbach and Geisse 2012). For rapid generation of material, transient expression in HEK293 or Chinese hamster ovary (CHO) cells is preferred.…”