Abstract：
:Fab's with hinges based on the human gamma1 sequence containing 1, 2, or 4 cysteines have been produced by high level Escherichia coli periplasmic secretion, and coupled in vitro by reduction/oxidation to form F(ab')2. We find that the F(ab')2 made with hinges containing 2 or 4 cysteines have a high level (approximately 70%) of multiple disulphide bonds. These F(ab')2 molecules have an increased pharmacokinetic stability as measured by area under the curve compared to those made by direct coupling through a single disulphide bond. One particular molecule containing 4 hinge cysteines has a greater pharmacokinetic stability than a F(ab')2 formed by chemical cross-linking. F(ab')2 made from the Fab' with 4 hinge cysteines is also relatively resistant to chemical reduction in vitro allowing partial reduction to expose reactive hinge thiols. These hinge sequences provide a simple method for producing robust F(ab')2 in vitro, obviating the need to use chemical cross-linkers, and provide a route to hinge specific chemical modification with thiol-reactive conjugates.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Humphreys DP,Vetterlein OM,Chapman AP,King DJ,Antoniw P,Suitters AJ,Reeks DG,Parton TA,King LM,Smith BJ,Lang V,Stephens PEdoi
10.1016/s0022-1759(98)00061-1subject
Has Abstractpub_date
1998-08-01 00:00:00pages
1-10issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(98)00061-1journal_volume
217pub_type
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