New developments in the production and use of stereoselective antibodies.
This article describes the production of stereoselective antibodies using both classical immunological and modern molecular biological techniques. Stereoselective antibodies against alpha-hydroxy acids were raised in rabbits and mice and compared with previously produced anti-alpha-amino acid antibodies. It was found that both types of antibodies combine stereoselectivity with class-specificity. Sequence analyses revealed that antibodies with opposing stereoselectivities can be formed during the affinity maturation process from a common progenitor or independently using nonhomologous binding sites. For the first time, phage display was employed to obtain stereoselective antibody fragments. The versatility of stereoselective antibodies as chiral selectors was demonstrated by applying them in several immunosensors and in chiral chromatography. A simple, membrane-based optical sensor allowed detection of enantiomeric impurities at the 1/2,000 level (99.9% ee). Silica-based antibody chiral stationary phases could be used for enantiomer separation of aliphatic amino acids in standard-sized columns, while miniaturized columns allowed interfacing with an MS-detector.
Hofstetter
Heike
H
Department of Chemistry and Biochemistry, Northern Illinois University, DeKalb, Illinois 60115-2862, USA.
Cary
James R
JR
Eleniste
Pierre P
PP
Hertweck
Jay K
JK
Lindstrom
Heather J
HJ
Ranieri
Daniel I
DI
Smith
Gregory B
GB
Undesser
L Pauline
LP
Zeleke
Jessica M
JM
Zeleke
Tigabu K
TK
Hofstetter
Oliver
O
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000276:immunology; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D002846:Chromatography, Affinity / Q000379:methods; D006880:Hydroxy Acids / Q000737:chemistry* / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D013237:Stereoisomerism
D000596:Amino Acids; D006880:Hydroxy Acids; D007135:Immunoglobulin Variable Region; D019151:Peptide Library