Antibody CR1-2B11 recognizes a non-polymorphic epitope of human CR1 (CD35).
Measurement of erythrocyte [red blood cells (RBC)] complement receptor type 1 (CR1, CD35) has the potential to serve as a sensitive assessment of complement activation and immune complex clearance. All previously reported monoclonal antibodies (MoAb) to the extracellular region of CR1 recognize epitopes within the long homologous repeats (LHR) of CR1 and the epitopes for the most frequently used MoAbs are repeated at least twice per CR1 molecule. Furthermore, CR1 exhibits structural polymorphism characterized by a variable number of LHR per molecule. Thus, accurate enumeration of cell surface CR1 using currently available MoAb would require that the results be corrected for the number of antibody epitopes per CR1 molecule encoded by each individual's alleles. To obtain a MoAb to a non-polymorphic epitope on human CR1, hybridomas were generated from mice immunized with recombinant soluble CR1 (sCR1) and MoAb were screened for those that recognized the full-length extracellular domain but failed to bind to all four recombinant LHR fragments. A single antibody, CR1-2B11, was identified and was found to recognize an epitope located wholly within SCR29-30 of CR1, NH2-terminal to an elastase cleavage site. Like other CR1 MoAb, the CR1-2B11 epitope expression decreased on old erythrocytes compared to younger cells and CR1-2B11 did not identify a CR1 'stump' on RBC. Importantly, CR1-2B11 immunofluorescence did not change with storage or handling of RBC, unlike the apparent decrease in immunofluorescence observed with other MoAb. CR1-2B11 should be useful for the accurate enumeration of RBC CR1.
Chen
C-H
CH
Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Boston, MA, USA.
Ghiran
I
I
Beurskens
F J M
FJ
Weaver
G
G
Vincent
J A
JA
Nicholson-Weller
A
A
Klickstein
L B
LB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western / Q000379:methods; D016466:CHO Cells; D016922:Cellular Senescence / Q000276:immunology; D006224:Cricetinae; D003412:Cricetulus; D000939:Epitopes / Q000097:blood*; D004912:Erythrocytes / Q000276:immunology; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D051379:Mice; D017463:Receptors, Complement 3b / Q000097:blood / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; C509991:CR1 protein, human; D000939:Epitopes; D017463:Receptors, Complement 3b; D011994:Recombinant Proteins