Anti-HUMIRA Antibodies

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1 Anti-HUMIRA Antibodies AbD Serotec offers you recombinant monoclonal antibodies to the human antibody adalimumab (HUMIRA). Our ready-made anti-adalimumab antibodies are ideal for preclinical research and clinical development. Type 1 Anti-Adalimumab Antibodies The Type 1 anti-adalimumab antibodies inhibit the binding of the drug adalimumab to its target, TNFα, and therefore detect free drug. These antibodies are ideal for use in direct or indirect 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. Product codes: HCA202, HCA203, HCA204 & HCA205 Type 3 Anti-Adalimumab/TNF-α Antibodies The Type 3 antibodies specifically recognize the adalimumab/tnfα complex. These antibodies detect adalimumab only when it is bound to TNF-α. They do not bind to free adalimumab or TNF-α (figure 3). With these antibodies, PK assays can be built that avoid the bridging format. In addition, they allow detection of adalimumab bound to its target in serum as opposed to free adalimumab. Product codes: HCA206, HCA207, HCA231 & HCA232 The anti-adalimumab antibodies are generated using HuCAL recombinant antibody technology, which ensures the continuity of supply and consistent performance needed for antibodies used in effective clinical trial assay development. The antibodies are sold for research use and come with an implied license for providing commercial in vitro research testing services. Anti-Adalimumab Antibodies Table 1 Antibody specifications Clone Product Code Immunogen Format Affinity kd, nm HCA202 Adalimumab Fab-FH Assay Development Recommendations Direct and indirect ELISA PK bridging ELISA with HCA204 Inhibition ELISA 18654_hIgG1 HCA203 Adalimumab Human IgG Inhibition ELISA Direct ELISA 18655_hIgG1 HCA204 Adalimumab Human IgG Inhibition ELISA PK bridging ELISA with HCA202 2 Direct ELISA 18664_hIgG1 HCA205 Adalimumab Human IgG HCA206 Adalimumab/TNF-α Fab-FH 1 67 PK ELISA assay, antigen capture format 18754_hIgG1 HCA207 Adalimumab/TNF-α Human IgG PK assay, antigen capture format HCA231 3 Adalimumab/TNF-α Fab-FH PK ELISA assay, antigen capture format 20349_hIgG1 HCA232 3 Adalimumab/TNF-α Human IgG PK assay, antigen capture format 1 FLAG - and His-6-tagged 2 Affinity measured in the monovalent Fab form 3 HCA231 and HCA232 are affinity matured versions of clone (HCA206 and HCA207, respectively)

2 Pharmacokinetic (PK) Assay - Bridging Format Product codes: HCA202 & HCA204 Anti-adalimumab antibody HCA202 was coated at 1.0 µg/ml on a microtiter plate and left over night. Washing and blocking was performed with 5% BSA in PBST. Adalimumab, titrated into 10% human serum, was then added. Detection was performed by adding HRP conjugated anti-adalimumab antibody HCA204* (2.0 µg/ml in HISPEC buffer, product code BUF049), plus QuantaBlu fluorogenic peroxidase substrate. The data shown in Figure 1 is the mean of three measurements. Figure 1 PK assay, bridging format, using antibodies HCA202 and HCA204 Inhibition ELISA Inhibition of adalimumab binding to TNF-α Product codes: HCA203 & HCA207 TNF-α (1.0 µg/ml) was used as the coating antigen followed by a pre-incubated mixture of the anti-idiotypic antibody HCA203 titrated into a constant amount of adalimumab (3.0 µg/ml). Free adalimumab, still capable of binding to the plate, was detected using HRP conjugated HCA207* (adalimumab/tnf-α complex specific). The data shown in Figure 2 is the mean of three measurements. Figure 2 Inhibition of adalimumab to TNF-α by antibody HCA203 * HCA204 & HCA207 were both conjugated to HRP using a LYNX Rapid Conjugation Kit (product codes LNK001P - LNK005P)

3 Antibodies Specificity Figure 3 Specificity of antibody HCA232 to drug-target complex Product code: HCA232 Figure 3 demonstrates the specificity of antibodies to the adalimumab/tnf-α complex. Type 3 antibodies (HCA206, HCA207, HCA231 and HCA232) will detect adalimumab only when bound to its target TNF-α (drug-target complex binders). These antibodies do not detect either adalimumab or TNF-α when they are not in a drug-target complex. Antigens were coated at 5.0 µg/ml on a microtiter plate overnight. After washing and blocking with 5% BSA, bound HRP-conjugated HCA232* was directly detected using QuantaBlu as substrate. Pharmacokinetic (PK) Assay Antigen Capture Format Figure 4 Antigen Capture ELISA using antibody HCA231 Product code: HCA231 Human TNF- was coated at 5 µg/ml on a microtiter plate. After washing and blocking with 5% BSA in PBST, adalimumab spiked into 10% human serum was added in the given concentrations. After washing, either unlabeled or HRP-labeled HCA231 were added at 2µg/mL in PBST. Unlabeled HCA231 was detected by adding HRP-labeled anti-his-antibody in HISPEC buffer (#BUF049C) and QuantaBlu fluorogenic peroxidase substrate. HRP-labeled HCA231 was detected directly with QuantaBlu substrate. Data are shown as the mean of three measurements. * Antibodies were conjugated to HRP using a LYNX Rapid Conjugation Kit (product codes LNK001P - LNK005P)

4 Ordering Information Specificity Product Code Clone Quantity Format ADALIMUMAB HCA mg Purified ADALIMUMAB HCA _hIgG1 0.1 mg Purified ADALIMUMAB HCA _hIgG1 0.1 mg Purified ADALIMUMAB HCA _hIgG1 0.1 mg Purified ADALIMUMAB (DRUG/TARGET COMPLEX) HCA mg Purified ADALIMUMAB (DRUG/TARGET COMPLEX) HCA _hIgG1 0.1 mg Purified Additional Anti-Biotherapeutic Antibodies Specificity Product Code Clone Quantity Format ALEMTUZUMAB HCA mg Purified ALEMTUZUMAB HCA _hIgG1 0.1 mg Purified ALEMTUZUMAB HCA175P 16942_hIgG1 0.1 mg Purified ALEMTUZUMAB HCA mg Purified ALEMTUZUMAB HCA _hIgG1 0.1 mg Purified BEVACIZUMAB HCA mg Purified BEVACIZUMAB HCA _hIgG1 0.1 mg Purified BEVACIZUMAB HCA184P 16748_hIgG1 0.1 mg HRP BEVACIZUMAB HCA _hIgG1 0.1 mg Purified CETUXIMAB HCA mg Purified CETUXIMAB HCA _hIgG1 0.1 mg Purified CETUXIMAB HCA _hIgG1 0.1 mg Purified CETUXIMAB HCA _hIgG1 0.1 mg Purified CETUXIMAB HCA _hIgG1 0.1 mg Purified INFLIXIMAB HCA mg Purified INFLIXIMAB HCA _hIgG1 0.1 mg Purified INFLIXIMAB HCA mg Purified INFLIXIMAB HCA _hIgG1 0.1 mg Purified INFLIXIMAB HCA _hIgG1 0.1 mg Purified OMALIZUMAB HCA _hIgG1 0.1 mg Purified OMALIZUMAB HCA mg Purified OMALIZUMAB (DRUG/TARGET COMPLEX) HCA _hIgG1 0.1 mg Purified OMALIZUMAB (DRUG/TARGET COMPLEX) HCA mg Purified RITUXIMAB MCA2260 MB2 A4 0.2 mg Purified RITUXIMAB MCA2260F MB2 A4 0.1 mg FITC RITUXIMAB HCA mg Purified RITUXIMAB HCA _hIgG1 0.1 mg Purified RITUXIMAB HCA mg Purified RITUXIMAB HCA062A µg Purified RITUXIMAB HCA mg Purified RITUXIMAB HCA061A µg Purified TRASTUZUMAB HCA mg Purified TRASTUZUMAB HCA mg Purified TRASTUZUMAB HCA mg Purified TRASTUZUMAB HCA mg Purified TRASTUZUMAB HCA _hIgG1 0.1 mg Purified TRASTUZUMAB HCA _hIgG1 0.1 mg Purified USTEKINUMAB HCA mg Purified USTEKINUMAB HCA mg Purified USTEKINUMAB HCA _hIgG1 0.1 mg Purified USTEKINUMAB HCA _hIgG1 0.1 mg Purified

5 View our secondary antibodies & related reagents online Americas Europe Toll Free: (USA & Canada) Tel: Tel: Fax: Fax: sales.de@abdserotec.com sales.us@abdserotec.com Worldwide Tel: Fax: sales@abdserotec.com LIT.PSADA Copyright Bio-Rad Laboratories, Inc All rights reserved. Published by AbD Serotec, a Bio-Rad Company. Endeavour House, Langford Lane, Kidlington, OXON, OX5 1GE. For in vitro research purposes and for commercial applications for the provision of in vitro research testing services. Any re-sale or any other form of commercial application needs a written agreement with AbD Serotec.HUMIRA is a registered trademark of Abbott Laboratories. HuCAL is a registered trademark of MorphoSys AG. QuantaBlu is a trademark of Thermo Fisher Scientific. LYNX Rapid Conjugation Kit is a registered trademark of Bio-Rad AbD Serotec Ltd. FLAG is a registered trademark of Sigma-Aldrich Co. LLC.

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