Download Intracellular Antibodies: Development and Applications by Antonino Cattaneo, Silvia Biocca PDF

By Antonino Cattaneo, Silvia Biocca

Recent advances within the box of recombinant antibodies have authorized the manipulation of genes encoding particular antibodies, therefore permitting their ectopic expression in a large choice of non-lymphoid cells. This quantity describes how the ectopic expression of antibodies, as secreted or as intracellularly retargeted molecules, might be exploited to dam organic features or to confer new phenotypic qualities (e.g. resistance to a virus). this is often the 1st publication describing this rising expertise, that's receiving expanding realization for software in lots of assorted fields and organic platforms - from human gene remedy to plant biotechnology.

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7): a. random mutations using mutator strains;69 b. targeted mutations;7o-7 2 c. chain shuffling;72, 73•74 d. error prone PCR;52, 61 ' 75 e. sexual PCR I DNA shuffling/6 When a selected phage antibody is expressed in a mutator strain, random mutations at the rate of approximately 1/2000 bp are inserted into the DNA. Some of these give rise to changes in antibody structure and hence affinity for the antigen under study. Passage of one or more specific phage antiborlies through a number of cycles of mutation in a mutator strain has been shown to increase the affinity of the derived phage antiborlies by 1oo-fold.

Chain shuffling;72, 73•74 d. error prone PCR;52, 61 ' 75 e. sexual PCR I DNA shuffling/6 When a selected phage antibody is expressed in a mutator strain, random mutations at the rate of approximately 1/2000 bp are inserted into the DNA. Some of these give rise to changes in antibody structure and hence affinity for the antigen under study. Passage of one or more specific phage antiborlies through a number of cycles of mutation in a mutator strain has been shown to increase the affinity of the derived phage antiborlies by 1oo-fold.

Kabat EA, Wu TT, Reid-Miller M et al. Sequences of proteins of immunological interest. US Department of Health and Human Services, US Government Printing Office 1987. 2. Chothia C, Lesk A. The relation between the divergence of sequence and structure in proteins EMBO J 1986; 5:823-826. 3· Chothia C, Lesk A. Canonical structures for the hypervariable regions of immunoglobulins. J. Biol Chem. 1987; 196:901-917. 4· Tramontana A, Chothia C, Lesk A. Framework residue 71 is a major determinant of the position and conformation of the second variable region in the VH domains of immunoglobulins.

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