Recombinant Antibody Production Service in Reticulocyte Cell-Free System

Cell-free systems utilizing rabbit reticulocytes have been established. Reticulocytes obtained from rabbits rendered anemic via acetylphenylhydrazine injection were processed into reticulocyte lysates. A method for the direct synthesis of target proteins without the background synthesis of endogenous proteins was developed using rabbit reticulocyte lysate. Endogenous mRNA was eliminated by subjecting the lysate to a micrococcal nuclease treatment. The relatively low protein output of systems for protein synthesis based on rabbit reticulocyte lysate is one of its limitations. According to a recent study, adding viral enhancers of translation can boost protein production by a factor of 10 or more.

Fig.1. Cell-free expression systems and their applications.¹

scFv-toxin fusion proteins were synthesized using rabbit reticulocyte lysate. In addition to E. Coli cell extracts, rabbit reticulocyte lysates were later employed as a fundamental translation system for the ribosome display-assisted in vitro evolution of antibody fragments. The most popular in vitro display methods are ribosome and mRNA display, which both allow the genotype and phenotype of a protein to be linked.

Advantages of Reticulocyte Cell-Free Systems

• Well-established system
• Mammalian system that allows for protein modification

Applications of Reticulocyte Cell-Free Systems

It has been demonstrated that peptide linkers with varying lengths and compositions can effectively connect the V_L and V_H domains of antibodies to form single-chain antibodies (sFv). A sFv may occasionally be a better choice for a therapeutic or diagnostic purpose than an equivalent mAb because of its unique properties. Proteins expressing in rabbit reticulocyte lysates that were a fusion of a sFv directed against TfnR and either the carboxyl-terminal 40 kDa of Pseudomonas exotoxin were discovered to be selectively cytotoxic to K562, a cell line that is known to express TfnR. This is the first time that functional recombinant antibodies have been generated in vitro.

Our commercially available cell-free systems, based on rabbit reticulocyte lysate, allow exogenous DNA to be transcribed and the resulting RNA to be translated in a single reaction. This eliminates the need to produce capped RNA before translation, which simplifies and lowers the cost of in vitro protein production.

Creative Biolabs is an expert in the field of recombinant antibody production. We have successfully developed and optimized the cell-free protein synthesis platform, and it has shown to be incredibly strong and effective for many challenging protein targets. Our experts have extensive experience designing the most dependable and economical plans to support our international clients' wide range of research initiatives. We provide recombinant antibody production services in reticulocyte cell-free systems to enable our clients worldwide to advance their varied research and projects. Please feel free to contact us for more details.

Reference

1. Tinafar, et al. "Synthetic biology goes cell-free." BMC biology 17 (2019): 1-14.

For research use only. Not intended for any clinical use.