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F(ab')2 or Fab' Preparation

  • General scheme for F(ab')2 preparation.
  • Example 1: Size exclusion HPLC analysis of F(ab')2 and Fab' produced at CellMosaic
Price:
Custom synthesis, please contact us for a quote.


In the preparation of antibody-enzyme conjugates there are a few advantages of using antibody fragments rather than intact antibodies. For example, antibody fragments lacking an Fc region have less interference with various Fc binding proteins, can easily penetrate cell membranes due to their lower molecule weight, and experience a lower nonspecific binding to cell surfaces. F(ab')2 and Fab' fragments are useful in immuno-histochemical studies. The procedure we use involves cleaving the IgG molecules below the disulfide groups in the hinge region using Pepsin or other suitable enzymes to create a bivalent fragment and degraded smaller peptide fragments, and then further reduction of purified F(ab')2 to produce Fab' fragment. 

Examples (see images):

  1. Synthesis and purification of F(ab')2 and Fab' fragments from IgG at CellMosaic (shown below): over 50% recovery for Fab' after purification. bm0007-fab-.jpg

References:

  1. Rousseaux, J.; Biserte, G.; Bazin, H. The differential enzyme sensitivity of rat immunoglobulin G subclasses to papain and pepsin. Molecular Immunology 1980, 17, 469-482.
  2. Coulter, A.; Harris, R. Simplified preparation of rabbit Fab fragments. J. Immun. Methods 1983, 59, 199-203.
  3. Rousseaux, R.; Rousseaux-Prevost, R.; Bazin, H. Optimal conditions for the preparation of Fab and F(ab')2 fragments from monoclonal IgG of different rat IgG subclasses. J. Immun. Methods 1983, 64, 141-146.

 

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<p>CellMosaic has Personalized Conjugation Kits (PerKit&trade;) for peptide labeling and conjugation, <a href="http://www.cellmosaic.com/personalized-conjugation-kits-perkit/">please click here to learn our&nbsp;PerKit&trade; product line</a>.</p>

<p>For large scale or project beyond the scope of&nbsp;PerKit&trade; configuration, <a href="http://www.cellmosaic.com/contact-us/">please contact us for a quote</a>.</p>

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