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Recombinant Antibodies

Recombinant Antibodies

Recombinant Antibodies

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.

Recombinant Antibodies

 

Recombinant Antibodies 2

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] (GTX635363) 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.

Iba1 antibody [HL22](GTX635363)


Figure 1. GeneTex's recombinant rabbit monoclonal Iba1 antibody [HL22] (GTX635363) is superior to a competitor's highly cited rabbit polyclonal antibody for both IHC-P (panels A vs. D) and IHC-Fr (panels B, C vs. E, F).

Another new rabbit recombinant product is GeneTex’s Ras (G12D mutant) antibody [HL10] (GTX635362), which specifically detects the G12D mutant by western blot with minimal background (Figure 2). Most importantly, it can also be used for IHC-P, where it demonstrates a robust signal and clear specificity for the G12D mutation in WT and KRAS G12D pancreatic tumor tissue sections (Figure 2A). This Ras (G12D mutant) antibody (GTX635362) is the first commercial recombinant version that shows exceptional specificity by IHC-P for this key tumorigenic mutant protein on sequence-verified human pancreatic tumor samples.

RAS (G12D Mutant) antibody [HL10](GTX635362)


Figure 2. (A) GeneTex's recombinant rabbit RAS (G12D mutant) antibody [HL10] (GTX635362) is sensitive and specific for the RAS G12D mutation by IHC of a KRAS G12D mutant pancreatic tumor tissue section (top) compared to a wild-type KRAS section (bottom). (B) The antibody is sensitive and specific for the RAS G12D mutation by WB of extracts from wild-type and mutant KRAS-confirmed human pancreatic cell lines. GAPDH for the loading control was detected by GTX100118. Lane1: HPDE. Lane2: HPNE. Lane3: AsPC1 (KRAS G12D). Lane4: BxPC3 (KRAS WT). Lane5: CFPAC1 (KRAS G12V). Lane6: HPAC (KRAS G12D). Lane7: HPAF-II (KRAS G12D). Lane8: MIA PaCa-22 (KRAS G12C). Lane9: PANC1 (KRAS G12D). Lane10: SU86.86 (KRAS G12D).

As mentioned above, the antigen-binding regions of a recombinant antibody can be inserted into various host IgG backbones or expressed in different binder formats (e.g., Fab fragments or scFvs). Here, GeneTex’s cloned TSG101 mouse monoclonal antibody (GTX70255) was converted to a rabbit IgG backbone (GTX635396) with preservation of performance. No cross-reaction was observed when the converted TSG101 rabbit IgG recombinant antibody was used in combination with an anti-mouse IgG secondary antibody (Figure 3).


Figure 3. Switch from a cloned mouse monoclonal antibody to a rabbit IgG backbone

 

Literature

  1. Brochure - Recombinant Antibodies
  2. Flyer - Recombinant Abs

 

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Flyer-Neuroscience

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Download the latest version of GeneTex's Recombinant Antibodies brochure.
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Brochure - Cited Antibodies

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Download the latest version of GeneTex's Cited Antibodies brochure.
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Flyer - Pancreatic Cancer Tumor Markers

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Poster - SARS-CoV-2 (COVID-19)

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Flyer - Cited Antibodies

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Protocols

Protocol - IHC (whole-mount)

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Protocol - Cytosolic-and Membrane Fractionation

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Single-Extracellular Vesicle Analysis (sEVA) to Detect Early Pancreatic Cancer

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Newsletter
PARPs as Therapeutic Targets in Clinical Oncology

PARPs as Therapeutic Targets in Clinical Oncology

Newsletter

Detection of VEEV Nonstructural Proteins with GeneTex’s Recombinant Monoclonal Antibodies

Newsletter
Article Alert: Natural Compounds with Broad Antiviral Activity against SARS-CoV-2 VOCs

Article Alert: Natural Compounds with Broad Antiviral Activity against SARS-CoV-2 VOCs

Newsletter
Influenza A and B Virus Recombinant Monoclonal Antibodies

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Influenza A and B Virus Recombinant Monoclonal Antibodies
Newsletter
The SARS-CoV-2 Omicron Variant

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HCK Inhibition: Boosting Immunotherapy’s Efficacy and Utility?

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Article Alert: C3aR and TNF-αR Antagonists as Potential Therapies for Medulloblastoma

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GeneTex’s “5 + 1” Pillar Antibody Validation Plan

Newsletter

Omicron BQ.1 and BF.7 Spike and Nucleocapsid Recombinant Proteins

Newsletter
New Recombinant Rabbit Interleukin-6 (IL-6) Monoclonal Antibody

New Recombinant Rabbit Interleukin-6 (IL-6) Monoclonal Antibody

Interleukin-6 (IL-6) is a cytokine with a high clinical profile due to its involvement in inflammatory processes, autoimmune disease, and cancer progression....
Newsletter

A Rising StAR in Cancer Research

Steroidogenic acute regulatory protein 1 (StAR/StARD1) mediates the rate-limiting step in steroidogenesis...
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Newsletter
SARS-CoV-2 can infect the human brain

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Newsletter
Nuclear PD-L1 Mediates Pyroptosis in Cancer Cells

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IHC- and FCM-validated Antibodies for SARS-CoV-2 Research

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Newsletter

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New Therapeutic Paradigm for Cancers with Activated NRF2

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Article alert: Proteomic Analysis Reveals How RNA Viruses Remodel Host Polysomes
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Skin Regeneration without Scar Formation

Skin Regeneration without Scar Formation

Skin Regeneration without Scar Formation
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mRNA Vaccine-induced Antibody Profiles against SARS-CoV-2 Variants

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mRNA Vaccine-induced Antibody Profiles against SARS-CoV-2 Variants
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GeneTex’s SARS-CoV-2 Spike RBD Antibodies Effectively Inhibit Viral Entry

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Neuropilins Facilitate SARS-CoV-2 Entry into Host Cells

Neuropilins Facilitate SARS-CoV-2 Entry into Host Cells
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Zika Virus Infection Increases Risk of Subsequent Severe Dengue Disease

Zika Virus Infection Increases Risk of Subsequent Severe Dengue Disease
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Spike Antibody-mediated Neutralization of SARS-CoV-2

Spike Antibody-mediated Neutralization of SARS-CoV-2

Spike Antibody-mediated Neutralization of SARS-CoV-2
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Profiling cytokine storm in COVID-19 patients by metatranscriptomic sequencing

Profiling cytokine storm in COVID-19 patients by metatranscriptomic sequencing

Profiling cytokine storm in COVID-19 patients by metatranscriptomic sequencing
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GeneTex Introduces Recombinant Monoclonal Iba1 Antibody

GeneTex Introduces Recombinant Monoclonal Iba1 Antibody

GeneTex's Iba1 recombinant antibody is suitable for IHC staining and WB.
Newsletter
GeneTex Launches Novel Recombinant RAS (G12D Mutant) Antibody [HL10]

GeneTex Launches Novel Recombinant RAS (G12D Mutant) Antibody [HL10]

GeneTex's RAS (G12D mutant) recombinant antibody is suitable for IHC staining and WB.
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Leaky blood-brain barrier triggers neural dysfunction that is reversible

Leaky blood-brain barrier triggers neural dysfunction that is reversible
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GeneTex Partners With BenchSci to Augment Antibody Catalog Publication Data

GeneTex Partners With BenchSci to Augment Antibody Catalog Publication Data

Partnership will use BenchSci's artificial intelligence to help researchers run successful experiments with the right antibody
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GeneTex Webinar Series

GeneTex Webinar Series
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SARS-CoV-2 Proteins and Antibodies for Lateral Flow-based Neutralization Assay

SARS-CoV-2 Proteins and Antibodies for Lateral Flow-based Neutralization Assay

for Lateral flow-based Neutralization Assay
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A Cancer Immunotherapy Holy Grail?

A Cancer Immunotherapy Holy Grail?
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MLLT3/AF9 is Pivotal for Human HSC Self-renewal and Engraftment

MLLT3/AF9 is Pivotal for Human HSC Self-renewal and Engraftment