Ideal for bioanalytical assays supporting preclinical research and clinical drug monitoring
Bio-Rad offers five ready made anti-idiotypic monoclonal antibodies to the chimeric rituximab (Rituxan®) antibody.
Anti-rituximab antibodies are ideal for use in direct ELISA, pharmacokinetic (PK) bridging assays and to develop and calibrate immune response (IR) assays to measure the anti-drug antibody (ADA) response in patient sera.
Antibody MCA2260, clone MB2 A4, was developed in rat using traditional antibody methods.
A further four antibodies were generated using the HuCAL® recombinant antibody library and phage display technology.
Antibody generation using HuCAL technology results in highly specific and sensitive antibodies, ideal for PK and PD assays. Antibodies are fully human, so are also perfect as controls or calibrators for ADA assays. The in vitro production of recombinant antibodies means that there is a consistent, secure supply throughout preclinical development and clinical trials, avoiding the need to develop new reagents.
More information about anti-idiotypic antibodies and their binding types and properties
|Product Code||Clone||Format||Affinity KD, nM||Assay Development Recommendations|
|HCA186||AbD18423||Monovalent Fab-FH1||0.13||PK bridging ELISA with MCA2660P as detection antibody|
|HCA201||AbD18423_hIgG1||Human IgG1||0.13*||ADA assay|
|MCA2260||MB2 A4||Rat IgG2a||Unknown||Direct ELISA|
|PK bridging ELISA with HCA186|
|Detection of rituximab in flow cytometry|
|PK bridging ELISA|
|PK bridging ELISA|
Table 1 Antibody Specifications
1 F=DYKDDDDK-tag H=His-6-tag;
2 Bivalent mini-antibody, M=c-myc-tag, H=His-6-tag);
* Affinity measured in the monovalent Fab format
Schematic image of PK Bridging ELISA. Anti-idiotypic capture antibody, Fab format (purple), monoclonal antibody drug (gold), anti-idiotypic detection antibody, Ig format (blue), labeled with HRP
Figure 1: PK assay, bridging format, using, antibodies HCA186 and MCA2260P
Figure 1: Anti-rituximab antibody, clone AbD18432 (HCA186), was coated on a microtiter plate at 1.0 µg/ml and left overnight. After washing and blocking with 5% BSA in PBST, rituximab spiked in 10% human serum was added in the given concentrations. Detection was performed by adding HRP conjugated anti-rituximab antibody, clone MB2A4 (MCA2260P), 3.0 µg/ml in HISPEC assay diluent (BUF049), plus QuantaBlu™ fluorogenic peroxidase substrate. Data are shown as the mean of three experiments.
Schematic image of ADA bridging assay. Monoclonal antibody drug as capture antibody and detection antibody labeled with HRP (gold), fully human anti-idiotypic antibody, Ig format (blue)
Figure 2: ADA assay, bridging, format using antibody HCA201
Figure 2: Rituximab was coated at two different concentrations (1.0 µg/ml and 1.5 µg/ml) on a microtiter plate and left over night. Anti-rituximab antibody, clone AbD18423_hIgG1 (HCA201), titrated into 10% human serum in the given concentrations, was then added. Detection was performed by adding HRP conjugated rituximab in HISPEC assay diluent, plus QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three experiments.
Figure 3: Specificity of anti-rituximab antibody HCA186
Figure 3: Demonstration of specificity to rituximab. Antigens (human serum as well as chimeric, humanized and human antibodies of the type IgG1/kappa) were coated at 5.0 µg/ml on a microtiter plate and left over night. After washing and blocking, anti-rituximab antibody, clone AbD18423 (HCA186) was added. Detection was performed using HRP conjugated mouse anti-His tag antibody (MCA5995P).
Licensed Use: For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
RITUXAN® is a registered trademark of Biogen Idec, Inc. USA. HuCAL® is a registered trademark of MorphoSys AG. QuantaBlu™ is a trademark of Thermo Fisher Scientific.