Anti-Rituximab (variable region) [MB2A4]

Absolute Antibody

Anti-Rituximab (variable region) [MB2A4]

9

This antibody is an idiotypic antibody against rituximab. It shows no cross-reactivity with other CD20-specific antibodies. Rituximab is a monoclonal antibody against CD20 used to treat autoimmune diseases and some types of cancer.

To develop an ELISA to measure rituximab, an ELISA was performed on solutions of rituximab using the rat version of this antibody. The ELISA could be used to quantify rituximab at concentrations between 0.125 and 50 microg/mL. Furthermore, the binding strength of this antibody to rituximab was measured using SPR. It showed a Kd value of 0.6 nM. Finally, a competition assay between this antibody and a P20 peptide was performed to determine whether P20 could block the binding of this antibody. A decrease in binding was indeed noted (Blasco et al., 2007; PMID: 17651747). To characterize this antibody, an ELISA was performed on diluted samples of rituximab using the rat version of this antibody. Furthermore, flow cytometry was used to detect rituximab on the surface of CD20-positive cells using this antibody. The antibody was bound to rituximab-coated cells and showed no cross-reactivity with other CD20-binding antibodies. The ability of this antibody to block rituximab from binding B cells was also shown (Cragg et al., 2004; PMID: 15213098). To determine the serum level of rituximab in pemphigus patients treated with rituximab, an ELISA was performed using the rat version of this antibody (Schmidt et al., 2009; PMID: 19502112). The rat version of this antibody was used in flow cytometry as an antibody that blocks rituximab from binding to B-cells to create an alternative pronase-free FCXM assay, as the presence of pronase can skew results of the normal assay (Alheim et al., 2018; PMID: 29157993). To determine the specificity of this antibody, flow cytometry was performed using the rat version of this antibody on CD20+ve Raji cells and CD20-ve K562 cells treated with rituximab. The antibody was shown to be highly specific for rituximab. Furthermore, an ELISA was performed on rituximab using this antibody (Hampson et al., 2010; PMID: 20547164). To determine the binding of rituximab to exosomes of B cells, a western blot was performed on exosomes incubated with rituximab using the rat version of this antibody. Furthermore, rituximab was detected in an ELISA using this antibody (Aung et al., 2011; PMID: 21873242). To study the structural alterations of rituximab in vivo, rituximab was immunoprecipitated from rat blood plasma using the rat version of this antibody. Furthermore, the plasma concentration of rituximab in rats was measured using ELISA with this antibody (Otani et al., 2017; PMID: 28052138). To prevent interference from rituximab, the rat version of this antibody was used to neutralize rituximab in a CDC-XM assay (Phillipe et al., 2013; PMID: 23994588).

Functional Assay, Flow Cytometry, ImmunoPrecipitation, Western Blot, ELISA, Neutralisation/Blocking, Other Application

Research Use Only

Monoclonal

MB2A4

Mouse

IgG2a

This chimeric mouse antibody was made using the variable domain sequences of the original Rat IgG2a format, for improved compatibility with existing reagents, assays and techniques.

Human

-20°C,2°C to 8°C

12352203