Synthetic antibody fragment as ligand in immunoaffinity chromatography.
The possibility that a fragment of an antibody molecule may interact with a protein antigen was tested by studying the binding properties of a thirteen-residue synthetic peptide with an amino acid sequence similar to part of a hypervariable segment of a monoclonal antibody directed against lysozyme. Affinity adsorbents were prepared with this peptide and with non-related peptides as ligand. Non-specific interactions could be abolished by washing the column with 0.05 M sodium thiocyanate in 20 mM tris-HCl (pH 7.4). Lysozyme was only bound to the antilysozyme adsorbent and could be eluted with 1 M sodium thiocyanate. The results show that immunoaffinity chromatography with synthetic peptide ligands which mimic the antigen-binding site may be a useful tool in the selective purification of proteins.
Welling
G W
GW
Laboratorium voor Medische Microbiologie, Rijksuniversiteit, Groningen, The Netherlands.
Geurts
T
T
van Gorkum
J
J
Damhof
R A
RA
Drijfhout
J W
JW
Bloemhoff
W
W
Welling-Wester
S
S
D000595:Amino Acid Sequence; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D008969:Molecular Sequence Data; D009113:Muramidase / Q000276:immunology
D000936:Antigen-Antibody Complex; D007128:Immunoglobulin Fragments; D009113:Muramidase