The production of biologics often involves cell culture or tissue culture, in which fetal bovine serum is widely used to provide essential nutrients for cell growth, and fetal bovine serum contains a certain amount of BSA (Bovine serum albumin). Considering the safety of the drugs produced, serum-free media are gradually used to replace serum-containing media in the production of biologics such as vaccines, antibody drugs, tissue-engineered products, and cells & GCTs, so as to minimize potential hazards such as viral contamination that may be brought about by serum. Among them, serum-free culture media are mostly formulated with various biomolecules that can replace the function of serum, such as bovine serum albumin (BSA), transferrin and insulin. As a result, BSA is present in both serum-containing and serum-free media, and although most of the impurity proteins are removed by purification and other steps in production, trace amounts of exogenous proteins (e.g., BSA, etc.) are still present in the final bioproducts.

BSA, as an exogenous protein, may cause serious allergic reactions after entering the human body, thus affecting the safety of biologics.The 2020 edition of the Chinese Pharmacopoeia (Part III) specifies the residual amount of BSA in various vaccines, which is basically no more than 50 ng/mL or no more than 50 ng/dose.

ELISA is a simple to use, sensitive and objective method for residue detection of BSA. This kit uses the principle of double-antibody sandwich enzyme-linked immunosorbent assay (sandwich ELISA) to detect the residual BSA in samples. First, add the BSA standard (36722-B) and test sample to the Anti-BSA coated microtiter strips (36722-A), and then diluted HRP-labeled BSA detecting antibody (36722-C) is added to form an antibody + antigen + antibody- HRP complex. HRP complex, wash the plate and add TMB color development mix (36722-E + 36722-F) to develop the color. TMB is catalyzed by HRP enzyme to change from colorless to blue and finally to yellow under the action of the termination solution (36722-G). The shade of yellow color is positively correlated with the amount of BSA detected in the sample. This kit can be used for the optimization of biologics purification processes, impurity control in intermediate processes, and final product release testing.

The detection range of this kit is 1.5625 ng/mL~50 ng/mL; the lower detection limit is ≤1ng/mL.

Features

l  Highly sensitive: Detecting as little as 1 ng/mL of BSA residuals in in-process and final biological samples.

l  Ensures accuracy: The BSA residuals can be accurately detected with a recovery of 75%-125%.

l  Specificity: Specific detection of BSA without interference from other exogenous proteins.

l  Stable: The difference between lot-to-lot is low, the kit property is not impacted under 37℃ for 7 days.

l  Easy signal collection: Using HRP-Biotin system, the signal can be stably enlarged.

Application

l  Residual BSA test in biological products

Specifications

Product Performance

Components

Storage

This product should be stored at 2°C ~8°C. Unopened product is valid for one year.

*Upon receipt of the kit, please check whether all components are complete and immediately store them in corresponding condition.

Figures

Precision Display

A) Repeatability

B) Intermediate precision

Table 1. The precision of BSA ELISA was assayed as the above scheme

10 duplicated were tested at 3 concentrations on the standard curve, and all the CV% were lower than 10% (Table 1A). 3 lot of BSA ELISA kits were tested and the CV% were lower than 10% (Table 1B).

Accuracy Validation

After the initial testing of BSA residue in four samples provided by the customer, the BSA content was measured to be within the range of the standard curve, and then the sample spiked recovery test was performed.

To the four samples to be tested (referred to as TS), 10ng/mL BSA standard was added in medium proportion, and the sample spiked recovery test was performed. The results showed that the spiked recoveries were all in the range of 80% to 120%.

Spiked recovery % = (measured value of spiked samples - measured value of unspiked samples)/theoretical value (amount of spiked standard) * 100%.

Table 2. Accuracy Validation

Documents:

Safety Data Sheet

36722_MSDS_HB250310.pdf

Manuals 

36722_Manual_Ver.EN20250310.pdf