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Antibodies for bioanalysis and drug monitoring of aflibercept and biosimilars

Develop highly selective and sensitive pharmacokinetic (PK) and anti-drug antibody (ADA) assays for aflibercept (Eylea) using our range of ready-made antibodies.

  • Inhibitory and non-inhibitory recombinant antibodies specific to aflibercept 
  • Antibody recognizing aflibercept VEGFR-Fc fusion region
  • Fully human surrogate positive control or calibrator
  • Sequence-defined, well-characterized reagents with a secure supply
  • Stringent quality control for batch-to-batch consistency
  • Non-animal-derived antibodies, reducing the use of animals in science

Table 1. Antibodies Specific to Aflibercept.

Specificity
Binding Type

Catalog Number

Clone

Format

Affinity* KD, nM

Assay Recommendation

Product Details

Aflibercept  Inhibitory

HCA386

AbD34644ia

hIgG1

11

PK bridging ELISA (detection)

Order HCA386

TZA0141P

AbD34644pap

Fab2-FH-X22-HRP2

11

PK bridging ELISA (detection)

Order TZA0141P

HCA387

AbD38645ia

hIgG1

1

ADA assay

Order HCA387

Aflibercept

Non-inhibitory

HCA389

AbD38720da

Fab-FSx21

1

PK bridging ELISA (capture)

Order HCA389

HCA390

AbD38778da

Fab-FSx21

4

PK bridging ELISA (capture)

Order HCA390

Aflibercept Fusion-region specific

Non-Inhibitory

HCA388

AbD38771ia

hIgG1

124

PK bridging ELISA (detection)

Order HCA388

TZA0142P

AbD38771pap

Fab2-FH-X22-HRP2

124

PK bridging ELISA (detection)

Order TZA0142P

* Affinity measured in the monovalent Fab format.
1Fab-FSx2 = Monovalent Fab antibody with (F) DYKDDDDK and (Sx2) StrepX-StrepX-tags.

Fab2-FH-X22-HRP = Bivalent Fab antibody with (F) DYKDDDDK, (H) His-6-tags, and Fab-SpyTag coupled to HRP-conjugated BiCatcher.


Anti-Aflibercept Antibodies

Aflibercept (Eylea, Zaltrap) is a recombinant fusion protein consisting of the extracellular domain of human VEGF receptors 1 and 2, fused to the Fc portion of human IgG1. It is used to treat the 'wet' form of age-related macular degeneration (AMD) and impaired vision due to macular oedema. It is also used in the treatment of metastatic colorectal cancer in combination with 5-fluorouracil, leucovorin and irinotecan in patients previously treated with an oxaliplatin-containing chemotherapy. Aflibercept binds to circulating VEGF-A and VEGF-B to inhibit their ability to promote angiogenesis. We offer inhibitory and non-inhibitory recombinant antibodies specific to aflibercept as well as an antibody recognizing aflibercept VEGFR-Fc fusion region.


Anti-Aflibercept Inhibitory Antibodies

Human Anti-Aflibercept Antibodies, clones AbD34644ia and AbD38645ia, are recombinant antibodies that specifically recognize the fusion protein drug aflibercept as well as VEGFR1. They do not recognize VEGFR2. The antibodies bind to free aflibercept and can be used to measure free aflibercept or aflibercept biosimilars in patient samples.

Clones AbD34644ia and AbD34644pap can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. It is recommended as the detection antibody paired with the Human Anti-Aflibercept Antibody, clone AbD3877da (HCA390), as the capture reagent. Clone AbD38645ia is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a surrogate positive control or reference standard in an anti-drug antibody (ADA) assay.


Anti-Aflibercept Non-Inhibitory Antibodies

Human Anti-Aflibercept Antibody, clone AbD38720da (HCA389), is a recombinant antibody that specifically recognizes the fusion protein drug aflibercept as well as VEGFR2. It does not recognize VEGFR1. It does not inhibit the binding of aflibercept to its target, VEGF; it binds to both free aflibercept and aflibercept bound to its target VEGF and can be used to measure total aflibercept and its biosimilars in bioanalytical assays.

A pair of anti-aflibercept antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure total drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with the Human Anti-Aflibercept Antibodies, clones AbD38771ia (HCA388) and AbD38771pap (TZA142P), as the detection antibodies.


Anti-Aflibercept Fusion Region-Specific Antibody

Human Anti-Aflibercept Antibody, clone AbD38771ia is a recombinant antibody that specifically recognizes the fusion protein drug aflibercept at the region of the fusion of VEGFR with the Fc portion of human IgG1. It does not inhibit the binding of aflibercept to its target, VEGF; it binds to both free aflibercept and aflibercept bound to its target VEGF and can be used to measure total aflibercept with high specificity. Clone AbD38771ia is a fully human recombinant monoclonal antibody with IgG1 isotype that can be used to develop a pharmacokinetic (PK) bridging assay to measure total drug. It is recommended as the detection antibody paired with the Human Anti-Aflibercept Antibody, clone AbD38720da (HCA389) as the capture reagent.


ELISA Protocols to Get You Started

PK Bridging ELISA

Fig.1 Aflibercept PK bridging ELISA using antibodies using antibodies HCA390 and HCA386.

Fig. 1. Aflibercept PK bridging ELISA using antibodies using antibodies HCA390 and HCA386.

Schematic image of PK bridging ELISA measuring free drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Ig format (blue), labeled with HRP.

Schematic image of PK bridging ELISA measuring free drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Ig format (blue), labeled with HRP.

Fig. 2. Aflibercept PK bridging ELISA using antibodies HCA390 and TZA0141P.

Fig. 2. Aflibercept PK bridging ELISA using antibodies HCA390 and TZA0141P.    

Schematic image of PK bridging ELISA measuring free drug.

Schematic image of PK bridging ELISA measuring free drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Fab format (purple), labeled with HRP.

Fig. 3. Aflibercept PK bridging ELISA using antibodies using antibodies HCA389 and HCA388.

Fig. 3. Aflibercept PK bridging ELISA using antibodies using antibodies HCA389 and HCA388.

Schematic image of PK bridging ELISA measuring total drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Ig format (blue), labeled with HRP

Schematic image of PK bridging ELISA measuring total drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Ig format (blue), labeled with HRP.

Fig. 4. Aflibercept PK bridging ELISA using antibodies HCA390 and TZA142P.

Fig. 4. Aflibercept PK bridging ELISA using antibodies HCA390 and TZA142P.  

Schematic image of PK bridging ELISA measuring free drug.

Schematic image of PK bridging ELISA measuring free drug. Capture antibody, Fab format (purple), fusion protein drug (gold-green), detection antibody, Fab format (purple), labeled with HRP.


ADA Bridging ELISA

Fig. 5. Aflibercept ADA bridging ELISA using antibody HCA387

Fig. 5. Aflibercept ADA bridging ELISA using antibody HCA387.

Schematic image of ADA bridging assay. Fusion protein drug as capture and detection reagent labeled with HRP (gold-green), fully human anti-drug antibody, Ig format (blue).

Schematic image of ADA bridging assay. Fusion protein drug as capture and detection reagent labeled with HRP (gold-green), fully human anti-drug antibody, Ig format (blue).

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