Monoclonal antibodies recognizing single amino acid substitutions in hemoglobin
Four monoclonal antibodies (mAb) to non-human primate hemoglobin referred to as Cap-4, Cap-5, Rh-2, and Rh-4, and two mAb to human hemoglobin, referred to as H-1 and H-3 were isolated and were partially characterized. Binding studies with these mAb on a panel of hemoglobins and isolated ..cap alpha.. and ..beta.. globin chains revealed a unique reactivity pattern for each mAb. Amino acid sequence analysis of the antigens used to generate the binding data suggests that the specific recognition of certain hemoglobin antigens by each mAb is controlled by the presence of a particular amino acid at a specific position within the epitope. The use of synthetic peptides as antigens confirmed this observation for five of the mAb. No synthetic peptides were tested with the sixth mAb, Rh-2. The amino acids required for binding of mAb Cap-4, Cap-5, Rh-4, and Rh-2 to hemoglobin are alanine at ..beta..5, threonine at ..beta..13, glutamine at ..beta..125, and leucine at ..cap alpha..68. The non-human primate hemoglobin antibodies require a specific amino acid that is not present in human hemoglobin. The amino acid required for binding of Cap-4, Cap-5, and Rh-4 could arise by a single base change in the ..beta.. globin gene, whereas the amino acid required for Rh-2 binding could only occur if two base changes occurred. Thus these mAb are candidate probes for a somatic cell mutation assay on the basis of the detection of peripheral blood red cells that possess single amino acid substituted hemoglobin as a result of single base substitutions in the globin genes of precursor cells.
- Research Organization:
- Univ. of California, Livermore
- DOE Contract Number:
- W-7405-ENG-48
- OSTI ID:
- 5459254
- Journal Information:
- J. Immunol.; (United States), Vol. 136:11
- Country of Publication:
- United States
- Language:
- English
Similar Records
Expression of fully functional tetrameric human hemoglobin in Escherichia coli
Development of an immunoassay to detect benzene adducts in hemoglobin
Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
HEMOGLOBIN
AMINO ACID SEQUENCE
MONOCLONAL ANTIBODIES
DIAGNOSTIC USES
MUTATIONS
DIAGNOSIS
AMINO ACIDS
BIOCHEMICAL REACTION KINETICS
CELL FLOW SYSTEMS
ERYTHROCYTES
HYBRIDOMAS
MAN
PRIMATES
ANIMAL CELLS
ANIMALS
ANTIBODIES
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
GLOBIN
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
KINETICS
MAMMALS
MATERIALS
MOLECULAR STRUCTURE
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PIGMENTS
PORPHYRINS
PROTEINS
REACTION KINETICS
USES
VERTEBRATES
551000* - Physiological Systems
550600 - Medicine