A recombinant antibody (rAb) is generated from the cloned, in vitro-expressed heavy and light chains of a selected monoclonal antibody obtained through classical hybridoma methodology, display strategies alone or in combination, sorted single B cells from immunized animals, or by one of many other techniques. The fact that it is cloned means the rAb is defined by its primary sequence, which can be engineered to optimize affinity and be expressed in different binder formats. The numerous advantages of rAbs have been well-documented, with consistency of performance being of paramount significance.
GeneTex’s rAb protocol employs a multi-parameter FACS-based approach to isolate antigen-specific IgG+ memory B cells from an immunized animal, with subsequent cloning of the antibody variable-region genes into an IgG backbone and expression in mammalian cells. This protocol is very rapid and can be completed in weeks, and also affords the opportunity to identify antibodies with diverse capabilities in various applications. Importantly, it allows cloning of the heavy and light chains from the same B cell, thereby preserving natural pairing. And once cloned, the supply of a given rAb is inexhaustible with exceptional reproducibility.
Why Recombinant Antibodies?
One outstanding advantage of this protocol is that the suitability of the individual clones for desired applications can be tested during screening. For example, GeneTex’s Iba1 rabbit recombinant antibody [HL22] detects Iba1, a protein commonly used as an immunohistochemical marker of both quiescent and activated microglia. In its development and production, both paraffin- and frozen-IHC analyses (IHC-P, IHC-Fr, respectively) were conducted in the first screening process to identify the best clones for these applications, as shown below (Figure 1). The clones were compared to a highly cited, market-leading commercial antibody to gauge their performance.
This strategy provides us with valuable perspective on a clone’s market competitiveness during development. In addition, our recombinant antibody team has successfully expressed the antigen-binding regions of this antibody in the context of both mouse and rat IgG backbones, thus extending flexibility for multiple staining.
|Product Name||Article No|
|RAS (G12D Mutant) antibody [HL10]||GTX635362|
|PD-L1 antibody [HL1041]||GTX635975|
|Estrogen Receptor alpha antibody [HL1082]||GTX636203|
|Androgen Receptor (ARv7 Splice Variant) antibody [HL1240]||GTX636639|
|Product Name||Article No|
|Iba1 antibody [HL22]||GTX635363|
|AChE antibody [HL1102]||GTX636298|
|OLIG2 antibody [HL1072]||GTX636104|
|GAD65 + GAD67 antibody [HL1181]||GTX636487|