Ranibizumab ELISA Kit

Sample

Aqueous humour

Species Reactivity

Human

Detection Method

ELISA

Intended Use

Enzyme immunoassay for quantitative analysis of free Ranibizumab in aqueous humour samples.

Contents of Kit

1. Microtiter Plate: 1 × 12 × 8. Break apart strips. Microtiter plate with 12 rows each of 8 wells coated with reactant.
2. Standard A-E: 0.3 mL (each).
Standards A-E (10×)
Standard A: 300 ng/mL
Standard B: 100 ng/mL
Standard C: 30 ng/mL
Standard D: 10 ng/mL
Standard E: 0 ng/mL
Used for the standard curve and control. Contains ranibizumab, human serum and stabilizer, <0,1% NaN₃.
Standards are prepared concentrated (10×). They should be diluted with the dilution rate given in the "Reagent Preparation" before the test.
3. Controls: 0.3 mL (each). Control low and high levels (10×), Contains stabilizer <0.1% NaN₃.
Controls are prepared concentrated (10×). They should be diluted with the dilution rate given in the "Reagent Preparation" before the test.
Control concentrations are given in "Quality control certificate"
4. Assay Buffer: 2 × 50 mL. Ready to use. Blue coloured. Contains proteins, <0.1% NaN₃.
5. Conjugate: 1 × 12 mL. Horse radish peroxidase conjugated probe. Ready to use. Red coloured. Contains HRP conjugated probe, stabilizer and preservatives.
6. Substrate: 1 × 12 mL. TMB substrate solution. Ready to use. Contains 3,3′,5,5′- Tetramethylbenzidine (TMB).
7. Stop Buffer: 1 × 12 mL. TMB stop solution. Ready to use. 1N HCl.
8. Wash Buffer (20×) : 1 × 50 mL. Prepared concentrated (20×) and should be diluted with the dilution rate given in the "Reagent Preparation" before the test. Contains buffer with tween 20.
9. Foil: 2 × 1. Adhesive Foil. For covering microtiter plate during incubation

Storage

The kit is shipped at ambient temperature (10-30°C) and should be stored at 2-8°C for long term storage. Keep away from heat or direct sunlight. The strips of microtiter plate are stable up to the expiry date of the kit in the broken, but tightly closed bag when stored at 2-8°C.

Performance Characteristics

Spike Recovery: 85% - 115%

Precision

Intra-assay and inter-assay CVs <30%

Detection Limit

0.625 ng/mL

Sensitivity

The lowest detectable level (Lowest detection limit, LOD) that can be distinguished from the zero standard is 0.625 ng/mL.
Functional sensitivity (Limit of quantification-LOQ): 1 ng/mL.

General Description

Ranibizumab is a recombinant humanized IgG1 kappa isotype monoclonal antibody fragment designed for intraocular use. Ranibizumab binds to the receptor binding site of active forms of VEGF-A, including the biologically active, cleaved form of this molecule, VEGF110. The binding of ranibizumab to VEGF-A prevents the interaction of VEGF-A with its receptors (VEGFR1 and VEGFR2) on the surface of endothelial cells, reducing endothelial cell proliferation, vascular leakage, and new blood vessel formation.
Ranibizumab is a recombinant humanized IgG1 kappa isotype monoclonal antibody fragment designed for intraocular use. Ranibizumab is a VEGF-A antagonist that binds to and inhibits the biologic activity of active forms of human VEGF-A, including the cleaved form (VEGF110). VEGF-A has been shown to cause neovascularization (angiogenesis) and an increase in vascular permeability, which is thought to contribute to the progression of the neovascular form of age-related macular degeneration (AMD).
Therapeutic drug monitoring (TDM) is the clinical practice of measuring specific drugs at designated intervals to maintain a constant concentration in a patient's bloodstream, thereby optimizing individual dosage regimens. The indications for drug monitoring include efficacy, compliance, drug-drug interactions, toxicity avoidance, and therapy cessation monitoring. Additionally, TDM can help to identify problems with medication compliance among noncompliant patient cases.
Biologic medicinal products (biologics) have transformed treatment landscapes worldwide for patients with haematological or solid malignancies with the 21st century. Today, as data exclusivity periods of first wave biologics approach expiration/have expired, several biosimilar products (i.e., biologics that are considered to be similar in terms of quality, safety and efficacy to an approved 'reference' biologic) are being developed or have already been approved for human use.
Like all biologics, biosimilars are structurally complex proteins that are typically manufactured using genetically engineered animal, bacterial or plant cell culture systems. As a consequence of this molecular complexity and the proprietary nature of the manufacturing process, which will inevitably result in the use of different host cell lines and expression systems as well as related differences in manufacturing conditions, it is not possible to manufacture exact copies of a reference biologic.
When administered to patients, all therapeutic proteins have the potential to induce an unwanted immune response (i.e., to stimulate the formation of anti-drug antibodies [ADAs]). The impact of immune responses can range from no apparent effect to changes in pharmacokinetics, loss of effect and serious adverse events. Furthermore, the immunogenicity profile of a biologic can be significantly altered by even small differences in its manufacturing process that are accompanied by a change in product attributes, as well as differences in dosing schedules, administration routes or patient populations.