Hybridomas display

Polyclonal antibodies can react with many epitopes, whereas MAbs are restricted to one epitope on proteins that do not have repeating sequences.24 By definition, polyclonal immunoassays are generally much more sensitive but less specific than monoclonal assays. Bispecific or hybrid antibodies can be used to increase the affinity. Bispecific antibodies are formed by the fusion of two previously established hybridomas to produce antibodies displaying the binding characteristics of both of the antibodies in one molecule.25... 

Krebber, A., S. Bornhauser, J. Burmester, A. Honegger, J. Willuda, H.R. Bosshard, and A. Pluckthun, Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system. J Immunol Methods, 1997. 201(1) 35-55. 

Catalytic antibodies, like enzymes, must be isolated and purified to homogeneity before they can be studied. Initially this was done by using the hybridoma technique for isolation of monoclonal antibodies (Box 31-A). After induction of antibody formation by injecting a selected hapten into a mouse, large numbers of monoclonal antibodies had to be tested for catalytic activity. Even if several thousand different monoclonal antibodies were tested, only a few with catalytic properties could be found.1 Newer methods have incorporated recombinant DNA techniques (Box 31-A) and use of combinatorial libraries and phage display.) Incorporation of acidic or basic groups into the haptens used to induce antibody formation may yield antibodies capable of mimicking the acid-base catalysis employed by natural enzymes. 0... 

An artificial immune system was constructed based on phage display to make antibodies without using rodents and hybridoma technology.23,26,27 The gene III... 

Depending on the isotype (i.e. class/subclass and kind of light chain), immunoglobulin molecules will display a particular biological property and will require an appropriate method for purification. The identification of mAb isotypes generally employs the culture supernatants of hybridomas and commercially available kits for the specific immunoenzymatic assays. This knowledge about the specific isotype facilitates the selection of the purification process in the next step. 

Ames RS, Tornetta MA, McMillan LJ, Neutralizing murine monoclonal antibodies to human IL-5 isolated from hybridomas and a filamentous phage Fab display library, J. Immunol., 1545 6355-6364, 1995. 

E. Phage Display Antibodies An Alternative to Hybridoma Technology ... 

A typical affinity maturation project using yeast cell surface display is divided into several steps. First, the variable (VH and VL) domains of the parent hybridoma clone are identified. Second, the VH and VL genes are cloned into a display vector for expression as a scFv fragment on the cell surface. Third, the binding characteristics of the WT scFv are determined. Fourth, diverse variegated populations for selection are created by mutagenesis of the parent clone plasmid DNA. Fifth, clones with improved affinities are enriched from the mutant library after application of selective pressure. Sixth, selected clones are sequenced and... 

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