Anti-Palivizumab Antibodies

Antibodies for bioanalytical method development to measure palivizumab and biosimilar products

Ready-made recombinant monoclonal anti-idiotypic antibodies, highly specific for the humanized antibody drug palivizumab (Synagis®).

Type 1 anti-idiotypic antibody

Anti-Palivizumab Type 1 Antibodies

The Type 1 anti-palivizumab antibodies inhibit the binding of the drug palivizumab to its target, a highly conserved region on the F protein of respiratory syncytial virus (RSV), and therefore detect free drug. These antibodies are ideal for use in indirect ELISA, to develop pharmacokinetic (PK) bridging assays and as a reference standard for anti-drug antibody (ADA) assays.

Product codes: HCA261, HCA262, HCA262P

These recombinant monoclonal antibodies are generated using the HuCAL® antibody library and an improved proprietary method of phage display, which results in highly specific and sensitive reagents. These are ideal for PK and PD assays. Antibodies are fully human, so they 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

Anti-biotherapeutic antibody quality control and characterization


Antibodies Specific to Palivizumab

Product Code Clone Antibody Specificity Binding Type Format Affinity KD, nM Assay Development Recommendations
HCA261 AbD23967 Palivizumab Type 1 Fab-FH1 1.5
Indirect ELISA
PK bridging ELISA as capture and detection antibody
PK bridging ELISA with HCA262
HCA262 AbD23967_hIgG1 Palivizumab Type 1 Human IgG1 1.5*
Indirect ELISA
PK bridging ELISA with HCA261
ADA assay
HCA262P AbD23967_hIgG1 Palivizumab Type 1 Human IgG1
HRP labeled
1.5* Direct ELISA
PK bridging ELISA with HCA261

Table 1 Antibody Specifications
1F=DYKDDDDK-tag H=His-6-tag
*Affinity measured in the monovalent Fab form


Pharmacokinetic Assay – Bridging ELISA

Pharmacokinetic Assay – Bridging ELISA

 

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

Anti-palivizumab antibody pair in PK bridging ELISA

Figure 1: Palivizumab PK ELISA bridging format using antibodies HCA261 and HCA262

Human anti-palivizumab antibodies, product codes HCA261 and HCA262

Figure 1: Anti-palivizumab antibody, clone AbD23967 (HCA261) was coated on a microtiter plate at 1.0 µg/ml and left over night. Washing and blocking was performed with PBST + 5% BSA. 10% human serum was added and spiked with increasing concentrations of palivizumab. Detection was performed using HRP conjugated anti-palivizumab antibody clone AbD23967_hIgG1 (HCA262P) diluted to 2 µg/ml in HISPEC buffer (BUF049A), plus QuantaBlu™ fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.

Protocol: PK bridging ELISA for use with anti-palivizumab antibodies 


Anti-Drug Antibody Assay - Bridging ELISA

Anti-Drug Antibody Assay - Bridging ELISA

 

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)

Use of fully human anti-palivizumab as calibrator in ADA assay

Figure 2: ADA assay, bridging format using antibody HCA262

Human anti-palivizumab antibody, product code HCA262

Figure 2: Palivizumab was coated at 0.5 µg/ml on a microtiter plate overnight. After washing and blocking with PBST + 5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of the human anti-palivizumab antibody clone AbD23967_hIgG1 (HCA262). Detection was performed using HRP conjugated palivizumab at 2 µg/ml in HISPEC assay diluent and QuantaBlu fluorogenic peroxidase substrate. Data are shown as the mean of three measurements. HRP conjugation of palivizumab was performed using a LYNX Rapid HRP Antibody Conjugation Kit®.

Protocol: ADA bridging ELISA for use with anti-palivizumab antibody HCA262


Demonstration of Antibody Specificity

Demonstration of the specificity of antibody HCA261 to palivizumab

Figure 3: Demonstration of the specificity of antibody HCA261 to palivizumab

Human anti-palivizumab antibody, product code HCA261

Figure 3: 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 anti-palivizumab antibody (clone AbD23967, HCA261) at a concentration of 2 µg/ml followed by an HRP labeled anti-DYKDDDDK tag antibody in HISPEC diluent and QuantaBlu fluorogenic peroxidase substrate.


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.

HuCAL® is a registered trademark of MorphoSys AG. QuantaBlu™ is a trademark of Thermo FisherScientific. Synagis® is a registered trademark of the AstraZeneca group of companies.