Recombinant Antibodies in Basic Neuroscience Research

Abstract

Basic neuroscience research employs antibodies as key reagents to label, capture, and modulate the function of proteins of interest. Antibodies are immunoglobulin proteins. Recombinant antibodies are immunoglobulin proteins whose nucleic acid coding regions, or fragments thereof, have been cloned into expression plasmids that allow for unlimited production. Recombinant antibodies offer many advantages over conventional antibodies including their unambiguous identification and digital archiving via DNA sequencing, reliable expression, ease and reliable distribution as DNA sequences and as plasmids, and the opportunity for numerous forms of engineering to enhance their utility. Recombinant antibodies exist in many different forms, each of which offers potential advantages and disadvantages for neuroscience research applications. I provide an overview of recombinant antibodies and their development. Examples of their emerging use as valuable reagents in basic neuroscience research are also discussed. Many of these examples employ recombinant antibodies in innovative experimental approaches that cannot be pursued with conventional antibodies.

Development, Screening, and Validation of Camelid‐Derived Nanobodies for Neuroscience Research

“…Other advantages of nAbs are (1) that only one polypeptide domain needs to be cloned and expressed; (2) expression in bacteria is typically higher than that of antigen‐binding fragments from conventional antibodies; and (3) nAbs exhibit a high degree of stability as compared to other miniaturized antibody forms. As recombinant antibodies, they also have unambiguous molecular definition, can be effectively archived as plasmids or as DNA sequence, and offer the potential for engineering to enhance their antigen‐binding properties and incorporate epitope tags, site‐specific labeling sequences, or other fusion partners to enhance detection of, and or confer biological function to the nAb (see Current Protocols article: Trimmer, 2020).…”

Immunofluorescence Microscopy

“…Moreover, the regions of the antibody gene that encode the antigen‐binding domains can be spliced to Fc regions from different isotypes, subtypes, or even different species. Collectively, these advantages allow recombinant antibodies to be a stable source of high‐quality antibodies that can be easily manipulated for different experimental purposes (Trimmer, 2020).…”

Controlling ion channel function with renewable recombinant antibodies

“…For these reasons, monoclonal antibodies or mAbs offer enhanced attributes in terms of research reproducibility as renewable and homogeneous preparations of a single antibody. Recombinant mAbs offer numerous additional advantages including allowing for antibody engineering to generate forms of antibodies that do not exist in nature but that have substantial advantages over conventional antibodies (Trimmer, 2020). Here we will focus on ion channel modulation by renewable forms of antibodies, including mAbs, recombinant antibodies in their various engineered forms, and antibody mimetics.…”

“…While the use of recombinant antibodies in basic biomedical research has been more limited, it is expanding in part due to advances in recombinant DNA techniques that have greatly reduced the time and effort needed for their development and subsequent engineering. There is also a growing recognition of the substantial advantages of recombinant antibodies as research reagents, in conforming to best practices in research transparency, rigor and reproducibility (Bradbury & Pluckthun, 2015 a , b ; Trimmer, 2020). These include: (1) unambiguous identification and definition of recombinant antibodies as produced in expression systems via DNA sequencing of the expression plasmids; (2) more reliable and less variable expression; (3) easier and more reliable dissemination as DNA sequences and as plasmids; and (4) permanent and absolute archiving as DNA sequence.…”