Serum-Free BLV gp51 Antigen Purification for ELISA Diagnostics
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Solution Overview
Problem
Current methods for purifying the BLV gp51 antigen for ELISA assays are labor-intensive, expensive, and prone to contamination, leading to false positive results due to the use of bovine serum in cell culture media, which complicates the rapid and effective diagnosis of enzootic bovine leukemia.
Innovation Solution
The production of purified BLV gp51 antigen is achieved using culture media completely devoid of bovine serum, specifically utilizing media like HyQ SFM4MegaVir, HyQ PF-Vero, and Gibco OptiPRO SFM, followed by ultrafiltration, dialysis, and purification on a DEAE-Sepharose FE column, resulting in a highly pure antigen for use in ELISA assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If bovine serum is used in cell culture media to produce gp51 antigen, then the antigen production is supported by available nutrients and growth factors, but the antigen becomes contaminated with bovine antibodies leading to false positive results
Solution Approach 1:
The patent removes bovine serum from the cell culture media composition, extracting the harmful component that causes contamination. This is achieved by formulating media without FCS while maintaining essential nutrients and growth factors through alternative sources, thereby eliminating bovine antibody contamination in the gp51 antigen preparation.
2Reliability
If traditional purification methods (precipitation, extraction, centrifugation, chromatography) are used to purify gp51 antigen, then antigen purity is improved, but the process becomes labor-intensive and expensive
Solution Approach 1:
The patent converts the harmful effect of bovine serum contamination into a beneficial selective purification approach. By using media without bovine serum, the contamination issue is prevented at the source, and the purification process is simplified to primarily involve ultrafiltration and dialysis rather than complex chromatographic procedures, reducing both labor and cost.
3Reliability
If avian serum is used instead of bovine serum in cell culture media, then bovine antibody contamination is eliminated, but a large quantity of avian antigens must be removed through additional purification steps
Solution Approach 1:
The patent adopts a disposable approach by using serum-free media formulations that are specifically designed for single-use in cell culture. This eliminates the need for complex removal procedures of avian or other foreign serums, as the media contains no such contaminants to begin with, thereby saving time and simplifying the purification process.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach eliminates contamination and reduces the complexity of antigen purification, enabling accurate and efficient diagnostics of enzootic bovine leukemia by providing a reliable, highly pure gp51 antigen for ELISA assays.
Implementation Method 1
purification on a DEAE-Sepharose FE column
Implementation Method 2
followed by ultrafiltration, dialysis, and purification
Implementation Method 3
followed by ultrafiltration, dialysis, and purification
Data Source
AI summary
The subject of the present invention is a method of obtaining purified BLV gp51 antigen as well as a novel ELISA assay using said antigen. The present invention is useful in the diagnosis of enzootic leukaemia in cattle.