Glycerol diversifies phage repertoire selections and lowers non-specific phage absorption.
Using a semi-synthetic phage displayed antibody repertoire, isoform-specific and cross-reactive phage-antibodies to eukaryotic elongation factor 1A (eEF1A) have been selected. Enrichment of specific antibodies was found to depend on the presence of glycerol. Further selections against lactate dehydrogenase (LDH) revealed that the dominance of a phage-antibody clone to LDH was inhibited by glycerol, a notable feature for selection strategies where a broad variety of binding clones is desired. The impact of glycerol in distinct steps of the selection protocol was examined and glycerol found to affect certain antibody-antigen interactions. Furthermore, the nonspecific phage binding was lowered by three orders of magnitude at a 20% (v/v) glycerol concentration.
Institute of Molecular and Structural Biology, University of Aarhus, Aarhus C, Denmark. kjaer@biobase.dk
S
S
Kjaer
B
B
Stausbøl-Grøn
T
T
Wind
P
P
Ravn
K
K
Hobolt Jensen
L
L
Kahns
B F
BF
Clark
D000906:Antibodies / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D003090:Coliphages / Q000187:drug effects* / Q000235:genetics; D017931:DNA Primers; D005990:Glycerol / Q000494:pharmacology*
D000906:Antibodies; D017931:DNA Primers; D005990:Glycerol