Anti-Simponi Antibodies

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1 Anti-Simponi Antibodies AbD Serotec offers you recombinant monoclonal antibodies to the antibody drug golimumab (Simponi). Our ready-made anti-golimumab antibodies are ideal for preclinical research and clinical assay development. Type 1 Anti-Golimumab Antibodies The Type 1 anti-golimumab antibodies inhibit the binding of the drug golimumab to its target, human TNF-α, 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: HCA240 and HCA241 Type 2 Anti-Golimumab Antibodies The Type 2 anti-golimumab antibody does not inhibit the binding of golimumab to TNF-α and can therefore detect total drug free, partially bound and fully bound. These fully human antibodies can be used to measure the golimumab levels in patient sera. Product code: HCA242 and HCA243 Type 3 Anti-Golimumab/TNF-α Antibodies The Type 3 antibodies specifically recognize the golimumab/tnf-α complex. These antibodies detect golimumab only when it is bound to TNF-α. They do not bind to free golimumab or TNF-α (Figure 5). With these antibodies, PK assays can be built that avoid the bridging format. In addition, they allow detection of golimumab bound to its target in serum as opposed to free golimumab. Product codes: HCA244 and HCA245 The anti-golimumab 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-Golimumab Antibodies Table 1 Antibody specifications Product Assay Development Clone Immunogen Format Affinity K Code D, nm Recommendations HCA240 Golimumab Fab-FH 1 Best capture antibody; 2.3 Direct ELISA; PK bridging ELISA; 2 Direct ELISA; PK bridging ELISA; ADA 20692_hIgG1 HCA241 Golimumab higg1 1.9 bridging ELISA HCA242 Golimumab Fab-FH 1 40 Detection antibody for PK assay 20897_hIgG1 HCA243 Golimumab higg Detection antibody for PK assay HCA244 Golimumab/TNF-α Fab-FH 1 85 (on complex) _hIgG1 HCA245 Golimumab/TNF-α higg1 (on complex) 1 FLAG - and His-6-tagged 2 Affinity measured in the monovalent Fab form PK ELISA assay, antigen capture format PK ELISA assay, antigen capture format

2 Pharmacokinetic (PK) Assay - Bridging Format ELISA Product codes: HCA240 & HCA241 Figure 1 PK assay, bridging format, using antibodies HCA240 and HCA241 A microtiter plate was coated over night with human anti-golimumab (HCA240) 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 golimumab. Detection was performed using HRP-conjugated antigolimumab (HCA241)* 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. Pharmacokinetic (PK) Assay Antigen Capture Format Figure 2 PK assay in antigen capture format ELISA, using Type 3 antibody HCA244 Product code: HCA244 Human TNF-α was coated at 5 µg/ml on a microtiter plate. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of golimumab. Detection was performed by using anti-golimumab (HCA244) at a concentration of 2 µg/ml, followed by an HRP labeled anti-his tag antibody* in HISPEC diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements. * Antibodies were conjugated to HRP using a LYNX Rapid Conjugation Kit (product codes LNK001P - LNK006P)

3 Anti-Drug Antibody (ADA) Assay Bridging ELISA Format Product Code: HCA241 Fig 3 ADA assay development, using Type 1 anti-golimumab antibody HCA241 A microtiter plate was coated over night with golimumab 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 Type 1 anti-golimumab (HCA241). Detection was performed using HRP-conjugated golimumab*. Data are shown as the mean of three measurements. Inhibition ELISA Inhibition of golimumab binding to TNF-α Product codes: HCA240 & HCA242 Figure 4 Inhibition of golimumab to TNF-α by Type 1 antibody HCA240, but not by Type 2 antibody HCA242 A microtiter plate was coated over night with human TNF-alpha at a concentration of 1 µg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of golimumab (0.3 µg/ml) plus increasing concentrations of either Type 1 (HCA240) or Type 2 (HCA242) anti-golimumab was added. Free golimumab, still capable of binding to the human TNF-alpha coated plate, was detected using a mouse anti-human IgG (Fc specific) HRP labeled antibody (MCA647P) and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

4 Antibody Specificity Figure 5 Specificity of Type 3 antibody HCA245 to drug-target complex Product code: HCA245 Figure 5 demonstrates the specificity of antibodies to the golimumab/tnf-α complex. Type 3 antibodies (HCA244 and HCA245) will detect golimumab only when bound to its target TNF-α (drugtarget complex binders). These antibodies do not detect either golimumab 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 PBS+5% BSA, detection was performed using anti-tnf-alpha/golimumab (HCA245) at a concentration of 2 μg/ml followed by an HRP labeled anti-his tag antibody* in HISPEC diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. * Antibodies were conjugated to HRP using a LYNX Rapid Conjugation Kit (product codes LNK001P - LNK006P) Ordering Information Specificity Product Code Clone Quantity Format GOLIMUMAB HCA mg Purified GOLIMUMAB HCA _hIgG1 0.1 mg Purified GOLIMUMAB HCA mg Purified GOLIMUMAB HCA _hIgG1 0.1 mg Purified GOLIMUMAB (DRUG/TARGET COMPLEX) HCA mg Purified GOLIMUMAB (DRUG/TARGET COMPLEX) HCA _hIgG1 0.1 mg Purified Additional Anti-Biotherapeutic Antibodies 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 Continues to the next page

5 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 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.PSGOL 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.Simponi is a registered trademark of Centocor Ortho Biotech Inc. 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.