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CG072 Anti-CD20 Monoclonal 2H7

B-lymphocyte antigen CD20 or CD20 is an activated-glycosylated phosphoprotein expressed on the surface of all B-cells beginning at the pro-B phase (CD45R+, CD117+) and progressively increasing in concentration until maturity.[3]

In humans CD20 is encoded by the MS4A1 gene.[4][5]

This gene encodes a member of the membrane-spanning 4A gene family. Members of this nascent protein family are characterized by common structural features and similar intron/exon splice boundaries and display unique expression patterns among hematopoietic cells and nonlymphoid tissues. This gene encodes a B-lymphocyte surface molecule that plays a role in the development and differentiation of B-cells into plasma cells. This family member is localized to 11q12, among a cluster of family members. Alternative splicing of this gene results in two transcript variants that encode the same protein.[5]

CD20 is the target of the monoclonal antibodies (mAb) rituximab, obinutuzumab, Ibritumomab tiuxetan, and tositumomab, which are all active agents in the treatment of all B cell lymphomas and leukemias. It has been shown that inhibition of BCR signalling by ibrutinib affects the expression of CD20 and the efficacy of anti-CD20 antibodies. [17]

The anti-CD20 mAB Ofatumumab (Genmab) was approved by FDA in Oct 2009 for Chronic lymphocytic leukemia.

The anti-CD20 mAB obinutuzumab (Gazyva) was approved by FDA in November 2013 for Chronic lymphocytic leukemia.




Product name
Anti-CD20    

Tested applications 
Bcell depletion, Flow,  ELISA, WB,  IHC

Immunogen 
Native Cd20

Form 
Liquid

Storage instructions
 Heat stable , shipped at ambient  temp  Upon delivery aliquot and store in fridge , longterm storage at  -20°C.

Storage buffer
PBS, pH 7.2

Concentration
 100 µl at 1.0  mg/ml

Purification 
Purified via protein G 

Isotype
Mouse IgG2b 


 
 
 


Standard Package 
100ug 

100ug

$380.00



Reference

Polyak, M. J. and J. P. Deans (2002). “Alanine-170 and proline-172 are critical determinants for extracellular CD20 epitopes; heterogeneity in the fine specificity of CD20 monoclonal antibodies is defined by additional requirements imposed by both amino acid sequence and quaternary structure.” Blood 99(9): 3256-3262. PubMed

In vivo ablation of malignant B cells can be achieved using antibodies directed against the CD20 antigen. Fine specificity differences among CD20 monoclonal antibodies (mAbs) are assumed not to be a factor in determining their efficacy because evidence from antibody-blocking studies indicates limited epitope diversity with only 2 overlapping extracellular CD20 epitopes. However, in this report a high degree of heterogeneity among antihuman CD20 mAbs is demonstrated. Mutation of alanine and proline at positions 170 and 172 (AxP) (single-letter amino acid codes; x indicates the identical amino acid at the same position in the murine and human CD20 sequences) in human CD20 abrogated the binding of all CD20 mAbs tested. Introduction of AxP into the equivalent positions in the murine sequence, which is not otherwise recognized by antihuman CD20 mAbs, fully reconstituted the epitope recognized by B1, the prototypic anti-CD20 mAb. 2H7, a mAb previously thought to recognize the same epitope as B1, did not recognize the murine AxP mutant. Reconstitution of the 2H7 epitope was achieved with additional mutations replacing VDxxD in the murine sequence for INxxN (positions 162-166 in the human sequence). The integrity of the 2H7 epitope, unlike that of B1, further depends on the maintenance of CD20 in an oligomeric complex. The majority of 16 antihuman CD20 mAbs tested, including rituximab, bound to murine CD20 containing the AxP mutations. Heterogeneity in the fine specificity of these antibodies was indicated by marked differences in their ability to induce homotypic cellular aggregation and translocation of CD20 to a detergent-insoluble membrane compartment previously identified as lipid rafts.




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