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  • Protocol for production and expression of chimeric bovine-human monoclonal antibodies.

Protocol for production and expression of chimeric bovine-human monoclonal antibodies.

STAR protocols (2023-01-04)
Behnaz Heydarchi, Damian F J Purcell
ABSTRACT

We describe herein a protocol for production of chimeric bovine-human monoclonal antibodies (mAbs) from vaccinated cows. The genes of HIV-1-specific single B cells are amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned into human expression vectors, and expressed in human cell lines. This protocol provides an efficient step-by-step methodology to produce HIV-1 chimeric mAbs and could be widely adapted for other antigens. For complete details on the use and execution of this protocol, please refer to Heydarchi et al. (2022).1.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Monoclonal Anti-Bovine IgG antibody produced in mouse, clone BG-18, ascites fluid
Sigma-Aldrich
Horse Serum, Donor Herd, USA origin, Heat inactivated, sterile-filtered, suitable for cell culture