Method for isolation of protein complexes using affinity binding
OwnerTomas Jelinek, Arthur LaMarche +2inventors
InventorsTomas Jelinek, Arthur LaMarche, Shannon K. Boyd, Dominic Eisinger
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Abstract
A method for the isolation and purification of a polypeptide or protein of interest as part of an antigen-antibody complex using a spin column is disclosed, wherein such methods employ either an antibody specific ligand derived from protein A and a spin column containing a resin that binds said ligand, or a spin column containing a nickel-chelate resin capable of directly binding an antibody. Kits containing compositions and spin columns useful in the purification methods are also described.
Claims15
A method for isolating a polypeptide as a purified polypeptide-antibody complex from a sample containing the polypeptide comprising:
- (a) contacting an antibody capture affinity ligand (ACAL) with a sample containing a polypeptide to be recovered from said sample, in the presence of an antibody that binds to said polypeptide to be isolated, under conditions promoting said contacting, to form a complex of said polypeptide, antibody and ACAL;
- (b) introducing the complex formed in step (a) to a spin column comprising a resin that binds said ACAL;
- (c) centrifuging the column of (b);
- (d) optionally washing the column of (c) with lysis buffer;
- and
- (e) washing the column from (d) with elution buffer to elute the polypeptide-antibody complex;
- thereby recovering a purified polypeptide-antibody complex.
A method for isolating a polypeptide as a purified polypeptide-antibody complex from a sample containing the polypeptide comprising:
- (a) contacting said sample, containing a polypeptide to be recovered, with an antibody that binds said polypeptide, under conditions promoting said contacting, to form a polypeptide-antibody complex;
- (b) introducing the complex formed in step (a) to a spin column containing a nickel-chelate resin that binds said antibody;
- (c) centrifuging the column of (b);
- (d) optionally washing the column of (c) with lysis buffer;
- and
- (e) washing the column from (d) with elution buffer to elute the polypeptide-antibody complex;
- thereby recovering a purified polypeptide-antibody complex.
Dependencies are inferred from the claim text.