Rekom Biotech is committed to ensure the highest quality level in the design and production of raw material for the IVD manufacturing industry.
Rekom Biotech products are designed, developed, produced and distributed according to our Quality Management System that is certified by ISO 9001 and ISO 13485 standards. Rekom antigens are always produced according to Standard Operating Procedures (SOPs) and undergo rigorous quality controls in our laboratories. You can check our system policy here.
Rekom Biotech is authorised to work with genetic modified organisms (GMO), with the license number A/ES/19/I-22, issued by National Biosafety Commission.
Rekom Biotech is registered as a Innovative SME, valid until Oct 25th 2021.
Each lot is subjected to various analyses:
Concentration detection by spectrophotometry
The measurement of the protein concentration is performed with the theoretical extinction coefficient of the recombinant protein obtained from Gill and vonHippel, 1989.
For proteins which do not contain any Trp residues, experience shows that this could result in more than 10% error in the computed extinction coefficient. Therefore, we measure the protein concentration by using the colorimetric assay based on the interaction between Coomassie brilliant blue and the arginine and aromatic residues (Bradford Method) and its maximum absorption shifts from 470 nm to 595 nm (Bradford, 1976).
Purity determination by SDS-PAGE
Aggregates presence analysed by size-exclusion chromatography (SEC)
Maldi-TOF for protein identification
Immunological analyses in ELISA or Western Blot assays.
For futher information, take a look at our technical report "Tritation experiments"
In this plot, the optical density at 450/620 nm for positive (blue) and negative (gray) IgG sera are compared for each concentration of the antigen. An appropriate statistical test of significance for the comparison of means between both groups, the Welch's test, is employed. Eligible concentrations for the use of the antigen should present statistically significant differences between positive and negative sera. This happens when the intervals at the top do not overlap and, equivalently, when the p-value at the bottom is below 0.05. In the present figure, all p-values are below 0.05 and thus the intervals do not overlap. Therefore, any of the showed concentrations can be used to distinguish between positive and negative sera.
We conjugate our antigens with biotin as an internal quality control for our proteins. We perform a spectroscopic estimation with HABA (4´-hydroxyazobenzene-2-carboxylic acid) of the mole-to-mole ratio of biotin to protein, also a western blot assay with streptavidin (A), and several ELISA assays (indirect ELISA assay in streptavidin coated microtiter plates, capture ELISA assay with the biotinylated antigen as detector and double-antigen-sandwich ELISA assay (B)).
(A) Western Blot assay with streptavidin
(B) Double antigen sandwich ELISA assay
Peroxidase (HRP) conjugation.
We also conjugate our antigens with peroxidase as an internal quality control. We perform a double antigen sandwich ELISA assay (C) and a capture ELISA assay by using a commercial test (D).
(C) A double antigen sandwich ELISA assay (DAS) performed with positive and negative CMV IgM specimen sera pre-validated with the ELISA capture IgM VIDAS.
(D) A capture ELISA assay performed with two different dilutions of the Rekom pp150-HRP in a reference commercial test (CMV-IgM-eLA test PKS medac).