By Nina Strebe, Frank Breitling, Dieter Moosmayer, Bodo Brocks, Stefan Dübel (auth.), Roland Kontermann, Stefan Dübel (eds.)
Antibodies are fundamental instruments for learn, analysis, and treatment. Recombinant ways enable the amendment and development of approximately all antibody homes, comparable to affinity, valency, specificity, balance, serum half-life, effector services, and immunogenicity.
Antibody Engineering offers a accomplished toolbox protecting the well-established fundamentals but additionally many intriguing new strategies. The protocols mirror the most recent "hands on" wisdom of key laboratories during this nonetheless fast-moving box. rookies will enjoy the confirmed step by step protocols, which come with beneficial useful recommendation; skilled antibody engineers will savor the hot principles and ways. The e-book is a useful source for all these engaged in antibody learn and improvement.
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Additional resources for Antibody Engineering
1, or for the expression of the antibody in a variety of formats. All vectors contain a chloramphenicol resistance cassette (camr) and additionally a tetracycline resistance “stuffer” cassette (tetA and tetR; 2,101 bp), which will be replaced by the antibody gene (the tet cassette allows the monitoring of complete SfiI digested vector by plating of transformed DH5a cells on tetracycline plates). V. Schaefer et al. to one RNeasy spin column and continue according to the manufacturer’s instructions.
1988; Glockshuber et al. 1990) is an ideal starting point for all engineering efforts, from sensors (Backmann et al. 2005; Morfill et al. 2007) to therapeutic fusion proteins (Di Paolo et al. 2003), or imaging reagents (Adams et al. 1993) to multivalent and multispecific reagents (Plu¨ckthun and Pack 1997), just to name a few illustrative examples. , to low picomolar KD for scFv fragments (Zahnd et al. 2004; Luginbu¨hl et al. 2006). Finally, some natural antibodies may not be of sufficient stability, which can also be corrected by engineering (Wo¨rn and Plu¨ckthun 2001; Ewert et al.
Elsevier, Amsterdam, pp 497–509 Backmann N, Zahnd C, Huber F, Bietsch A, Plu¨ckthun A, Lang HP, Gu¨ntherodt HJ, Hegner M, Gerber C (2005) A label-free immunosensor array using single-chain antibody fragments. Proc Natl Acad Sci USA 102:14587–14592 Barbas C, Burton D, Scott J, Silverman G (2001) Phage display: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY Bothmann H, Plu¨ckthun A (1998) Selection for a periplasmic factor improving phage display and functional periplasmic expression.
Antibody Engineering by Nina Strebe, Frank Breitling, Dieter Moosmayer, Bodo Brocks, Stefan Dübel (auth.), Roland Kontermann, Stefan Dübel (eds.)