Catalog No.
KDB86902
Description
PRINCIPLE OF THE ASSAY This assay employs the quantitative competitive enzyme immunoassay technique. Recombinant Human EGFR has been pre-coated onto a microplate. Standards or samples are premixed with biotin-labeled antibody and then pipetted into the wells. Losatuxizumab in the sample competitively binds to the pre-coated protein with biotin-labeled Losatuxizumab. After washing away any unbound substances, Streptavidin-HRP is added to the wells. Following a wash to remove any unbound enzyme reagent, a substrate solution is added to the wells and color develops in inversely proportion to the amount of Losatuxizumab bound in the initial step. The color development is stopped and the intensity of the color is measured.
Applications
Used for the quantitative determination of Losatuxizumab concentration in serum and plasma.
Detection method
Colorimetric
Sample type
Plasma, Serum
Assay type
Quantitative
Range
234.38 - 15,000 ng/mL
Sensitivity
73.16 ng/mL
Precision
Intra-Assay Precision (Precision within an assay): <20%
Three samples of known concentration were tested sixteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays): <20%
Three samples of known concentration were tested in twenty four separate assays to assess inter-assay precision.
|
Intra-Assay Precision |
Inter-Assay Precision |
||||
Sample |
1 |
2 |
3 |
1 |
2 |
3 |
n |
16 |
16 |
16 |
24 |
24 |
24 |
Mean (ng/mL) |
10254.8 |
2063.8 |
583.1 |
6845.3 |
2308.1 |
604.8 |
Standard deviation |
1196.0 |
105.2 |
52.2 |
478.9 |
136.0 |
34.1 |
CV (%) |
11.7 |
5.1 |
8.9 |
7.0 |
5.9 |
5.6 |
Recovery
80-120%
Shipping
2-8 ℃
Stability and Storage
When the kit was stored at the recommended temperature for 6 months, the signal intensity decreased by less than 20%.
Alternative Names
ABT-806, PR-1316749, CAS: 1801544-27-3
Background
Losatuxizumab is a biosimilar that targets EGFR. EGFR (Epidermal growth factor receptor, HER1, ErbB1) is encoded by the EGFR gene located on chromosome 7 in humans. EGFR belongs to the HER/ERbB family of proteins that includes three other receptor tyrosine kinases, ERbB2, ERbB3, ERbB4. EGFR is a transmembrane receptor and binding of its cognate ligands such as EGF (Epidermal Growth Factor) and TGF alpha (Transforming Growth Factor alpha) to the extracellular domain leads to EGFR dimerization followed by autophosphorylation of the tyrosine residues in the cytoplasmic domain. Phosphorylation of EGFR at certain residues is also mediated by Src-non-receptor kinase. EGFR activation signals multiple downstream signaling cascades such as the Ras - ERK, pi3-K - Akt, Jak - STAT and PKC pathways that help in growth and proliferation of cells. Phosphorylation of EGFR at Y1086 specifically allows binding of the adaptor protein GRB2, leading to activation of the MAPK pathway. Upon receptor activation and signaling, EGFR is endocytosed and targeted for degradation or recycling. Mutations in the EGFR gene are associated with lung cancer and multiple alternatively spliced transcript variants encode different protein isoforms of EGFR have been found. Increased production or activation of EGFR has been associated with poor prognosis in a variety of tumors. Moreover, EGFR overexpression is observed in tumors of the head and neck, brain, bladder, stomach, breast, lung, endometrium, cervix, vulva, ovary, esophagus, stomach and in squamous cell carcinoma.