Vedolizumab antibody | AbD30136_hIgG1
A microtiter plate was coated over night with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 (HCA293) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with vedolizumab, human IgG1/kappa and human IgG1/lambda at a concentration of 5 µg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 (HCA293) titrated to the given concentrations in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data for vedolizumab are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA292) at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of vedolizumab. Detection was performed using HRP conjugated Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 (HCA293) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).
A microtiter plate was coated overnight with vedolizumab at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 (HCA293, black line), clone AbD30138_hIgG1 (HCA294, red line) or clone AbD30139_hIgG1 (HCA295, blue line). Detection was performed using HRP conjugated vedolizumab. HRP conjugation of the vedolizumab used in this assay was performed using a LYNX Rapid HRP Conjugation Kit® (LNK002P). Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with recombinant human α4β7 integrin at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of vedolizumab (0.3 µg/ml), spiked with increasing concentrations of Human Anti-Vedolizumab Antibody, clone AbD30136 (HCA293) in the monovalent Fab format was added. Free vedolizumab, still capable of binding to the human α4β7 integrin coated plate, was detected using a Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti Vedolizumab
Human Anti-Vedolizumab Antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in drug level monitoring and immunogenicity testing. It can be used as detection antibody in a PK bridging ELISA and as a calibrator in an anti-drug antibody assay.
- Product Type
- Monoclonal Antibody
- Clone
- AbD30136_hIgG1
- Isotype
- IgG1
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA293 | E | 0.1 mg |
Human Anti-Vedolizumab Antibody, clone AbD30136_hIgG1 is a paratope specific, inhibitory anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug vedolizumab. The antibody does not recognize free recombinant human α4β7 integrin or vedolizumab in complex with α4β7 integrin and can be used to measure free vedolizumab levels in serum from patients.
Clone AbD30136_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay. A pair of anti-vedolizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody paired with clone AbD30136 (HCA292) in monovalent Fab format as the capture antibody.
Vedolizumab (Entyvio) is a humanized antibody (IgG1/kappa), derived from a murine antibody. It is used for the treatment of both moderate to severe ulcerative colitis and moderate to severe Crohn's disease, primarily in patients who have lost response to, or are intolerant to inhibitors of tumor necrosis factor alpha (TNFα), such as infliximab, adalimumab. The drug binds specifically to human α4β7 integrin, a key mediator of gastrointestinal inflammation expressed on the surfaces of T and B lymphocytes. By selectively inhibiting the binding of α4β7 integrin to the mucosal addressin cell adhesion molecule-1 (MAdCAM-1), vedolizumab prevents leukocyte binding to the endothelial surface and extravasation into affected tissue. Specifically inhibiting this pathway alleviates gastrointestinal inflammation without impairing systemic immune responses.
View a summary of all anti-vedolizumab antibodies
Clone AbD30136_hIgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay. A pair of anti-vedolizumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody paired with clone AbD30136 (HCA292) in monovalent Fab format as the capture antibody.
Vedolizumab (Entyvio) is a humanized antibody (IgG1/kappa), derived from a murine antibody. It is used for the treatment of both moderate to severe ulcerative colitis and moderate to severe Crohn's disease, primarily in patients who have lost response to, or are intolerant to inhibitors of tumor necrosis factor alpha (TNFα), such as infliximab, adalimumab. The drug binds specifically to human α4β7 integrin, a key mediator of gastrointestinal inflammation expressed on the surfaces of T and B lymphocytes. By selectively inhibiting the binding of α4β7 integrin to the mucosal addressin cell adhesion molecule-1 (MAdCAM-1), vedolizumab prevents leukocyte binding to the endothelial surface and extravasation into affected tissue. Specifically inhibiting this pathway alleviates gastrointestinal inflammation without impairing systemic immune responses.
View a summary of all anti-vedolizumab antibodies
Product Details
- Product Form
- Human IgG1 antibody (lambda light chain) selected from the HuCAL® phage display library and expressed in a human cell line. This antibody is supplied as a liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Vedolizumab
- Affinity
- The monovalent intrinsic affinity of AbD30136_hIgG1 was measured as KD= 0.1 nM by real time, label free molecular interaction analysis on immobilized vedolizumab.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
More Information
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany. Entyvio is trademark of Millennium Pharmaceuticals, Inc.
- 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.
- Regulatory
- For research purposes only
Applications of Vedolizumab antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Technical Advice
- Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- Clone AbD30136_hIgG1 can be used in a direct or indirect ELISA or as detection antibody for vedolizumab in a bridging ELISA together with (HCA292) as the capture reagent. Furthermore, it may be used as a calibrator for the set-up of an anti-drug antibody assay.
Protocol: PK bridging ELISA to measure free drug and ADA bridging ELISA.
Copyright © 2019 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | 50 ml | E IY | ||
| Human anti Vedolizumab | HCA292 | 0.1 mg | E | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | 3 Conjugations For 400µg Antibody | CJ | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | 0.2 mg | C* E |
