Introduction to Membrane Protein Antibody Discovery and Phage Display

The preparation of membrane protein immunogens has been a difficult task in membrane protein antibody production due to the difficulty of maintaining their activity and natural conformation in vitro. Creative Biolabs provides an advanced and well-established platform for membrane protein research that can facilitate the discovery and production of your membrane protein antibodies using phage display technology.

What Is Phage Display?

Phage display technology is an efficient gene expression screening technique. It involves the fusion of an exogenous protein or peptide with a specific phage capsid protein, which is then displayed on the phage surface and maintained in a relatively independent spatial conformation and biological activity. This facilitates the specific recognition and binding of the target molecule, thus achieving the unification of genotype and expression type.

Specifically, phage display technology involves the insertion of exogenous genes encoding polypeptides or proteins into the appropriate positions of phage shell protein structural genes by genetic engineering techniques. The objective of selecting the appropriate position is to enable correct expression in the reading frame so that the exogenous polypeptide or protein forms a fusion protein on the coat protein of the phage. The fusion protein is presented on the surface of the phage with the reassembly of the zygotic phage. At this point, the protein can maintain its relative spatial structure and biological activity. The target molecule is then used to wash away the unspecifically bound phage using a suitable elution method. The bound phage is then eluted with acid-base or competing molecules, and the neutralized phage is infected with E. coli for amplification, and after 3-5 rounds of enrichment, the proportion of phage that can specifically recognize the target molecule is gradually increased, and finally, a peptide or protein that recognizes the target molecule is obtained.

Membrane Protein Antibodies and Phage Display

Phage display technology provides a simple and rapid route to genetically engineered antibody preparation for the production of monoclonal antibodies. Antibody screening using phage display technology can combine selection and amplification capabilities with powerful screening capabilities that can mimic the in vitro process of membrane protein antibody production in vivo, allowing the preparation and production of membrane protein antibodies without immunization. This greatly simplifies the process of membrane protein antibody production and greatly increases the success rate of antibody production.

The phage display technique for constructing antibody libraries eliminates the cell fusion step, avoids the tedious procedure of repeated subcloning due to the instability of hybridomas, and greatly increases the library capacity from several thousand clones of hybridomas up to 106. Phage display technology can directly obtain antibody genes, which can facilitate the further construction of various genetically engineered antibodies, and can also be used for some difficult-to-prepare antibodies, such as weak immunogens and toxic antigens, as well as humanized antibodies. The short cycle time of phage display technology and the value added in bacteria make it suitable for large-scale industrial production of membrane protein antibodies.

Creative Biolabs would like to share with you the key points and experiences in membrane protein antibody discovery and phage display technology to facilitate the smooth running of your project.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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