Epitope-directed monoclonal antibody production using a mixed antigen cocktail facilitates antibody characterization and validation

AbstractHigh quality, well-validated antibodies are needed to mitigate irreproducibility and clarify conflicting data in science. We describe an epitope-directed monoclonal antibody (mAb) production method that addresses issues of antibody quality, validation and utility. The workflow is illustrated by generating mAbs against multiple in silico-predicted epitopes on human ankyrin repeat domain 1 (hANKRD1) in a single hybridoma production cycle. Antigenic peptides (13–24 residues long) presented as three-copy inserts on the surface exposed loop of a thioredoxin carrier produced high affinity mAbs that are reactive to native and denatured hANKRD1. ELISA assay miniaturization afforded by novel DEXT microplates allowed rapid hybridoma screening with concomitant epitope identification. Antibodies against spatially distant sites on hANKRD1 facilitated validation schemes applicable to two-site ELISA, western blotting and immunocytochemistry. The use of short antigenic peptides of known sequence facilitated direct epitope mapping crucial for antibody characterization. This robust method motivates its ready adoption for other protein targets.

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A Call for a European Prohibition of Monoclonal Antibody Production by the Ascites Procedure in Laboratory Animals

Alternatives to Laboratory Animals ◽

10.1177/026119299802600414 ◽

1998 ◽

Vol 26 (4) ◽

pp. 523-536

Author(s):

Coenraad F.M. Hendriksen

Keyword(s):

Monoclonal Antibody ◽

Laboratory Animals ◽

Induction Method ◽

Therapeutic Application ◽

Monoclonal Antibody Production ◽

In Vitro Methods ◽

Appeals Process ◽

Core Facilities ◽

Mab Production

Monoclonal antibodies (mAbs) are particularly valuable in therapeutics and research. Unfortunately, one of the most familiar methods of producing mAbs, the ascites induction method, causes pain and distress to the animals used. In most cases, non-animal or in vitro alternatives can be employed to reduce or eliminate the use of animals for mAb production. Prohibition of the use of animals in the production of mAbs is recommended, except when the replacement in vitro methods prove to be insufficient, and in a limited number of other well-documented cases, such as an exceptional need for an emergency therapeutic application. A total ban on the use of animals for mAb production is impractical and it is imperative that an appeals process should accompany the prohibition. The need for the establishment of core facilities for in vitro mAb production is emphasised.

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Detailed Review of Monoclonal Antibody Production and Its Advantage as a Role in Malignancy and Transplant Rejection

International Journal of Scientific and Research Publications (IJSRP) ◽

10.29322/ijsrp.8.11.2018.p8337 ◽

2018 ◽

Vol 8 (11) ◽

Author(s):

Chandrasekaran M M. ◽

N. Sivakumaran

Keyword(s):

Monoclonal Antibody ◽

Antibody Production ◽

Transplant Rejection ◽

Monoclonal Antibody Production ◽

Detailed Review

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Faculty Opinions recommendation of The ankyrin repeat domain of the TRPA protein painless is important for thermal nociception but not mechanical nociception.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.13755956.15175057 ◽

2012 ◽

Author(s):

Bernd Nilius

Keyword(s):

Ankyrin Repeat ◽

Thermal Nociception ◽

Repeat Domain ◽

Ankyrin Repeat Domain

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Asparaginyl Hydroxylation of the Notch Ankyrin Repeat Domain by Factor Inhibiting Hypoxia-inducible Factor

Journal of Biological Chemistry ◽

10.1074/jbc.m704102200 ◽

2007 ◽

Vol 282 (33) ◽

pp. 24027-24038 ◽

Cited By ~ 147

Author(s):

Mathew L. Coleman ◽

Michael A. McDonough ◽

Kirsty S. Hewitson ◽

Charlotte Coles ◽

Jasmin Mecinović ◽

...

Keyword(s):

Hypoxia Inducible Factor ◽

Ankyrin Repeat ◽

Repeat Domain ◽

Ankyrin Repeat Domain

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Preliminary Techno-Economic Assessment of Animal Cell-Based Meat

Foods ◽

10.3390/foods10010003 ◽

2020 ◽

Vol 10 (1) ◽

pp. 3

Author(s):

Derrick Risner ◽

Fangzhou Li ◽

Jason S. Fell ◽

Sara A. Pace ◽

Justin B. Siegel ◽

...

Keyword(s):

Monoclonal Antibody ◽

Chemical Engineering ◽

Animal Cell ◽

Economic Assessment ◽

The United States ◽

Economic Viability ◽

Monoclonal Antibody Production ◽

Technological Performance ◽

Low Volume ◽

Cost Factors

Interest in animal cell-based meat (ACBM) or laboratory-grown meat has been increasing; however, the economic viability of these potential products has not been thoroughly vetted. Recent studies suggest monoclonal antibody production technology can be adapted for the industrialization of ACBM production. This study provides a scenario-based assessment of the projected cost per kilogram of ACBM produced in the United States based on cellular metabolic requirements and process/chemical engineering conventions. A sensitivity analysis of the model identified the nine most influential cost factors for ACBM production out of 67 initial parameters. The results indicate that technological performance will need to approach technical limits for ACBM to achieve profitably as a commodity. However, the model also suggests that low-volume high-value specialty products could be viable based on current technology.

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