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Published on: November 21, 2018
High-throughput 96-well plate-based porcine antibody isolation protocol
John Byrne1, Christina Bourne1, Sitka Eguiluz1
1Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, United States of America.
Scientists developed a high-throughput 96-well plate method for isolating swine immunoglobulin (Ig) isotypes, enabling efficient measurement of influenza-specific antibodies in pigs. This technique accelerates antibody isolation from various samples, crucial for immunogenicity studies.
Area of Science:
- Immunology
- Biochemistry
- Analytical Chemistry
Background:
- Column chromatography is a standard technique for analyte purification and antibody isolation.
- Existing methods struggle to detect specific antibodies due to physiological variations and sample complexities.
- High background components in swine serum, milk, and colostrum complicate antibody detection.
Purpose of the Study:
- To develop a high-throughput antibody isolation protocol for measuring influenza-specific antibodies in pigs.
- To overcome challenges associated with high background in swine biological fluids.
- To enable efficient analysis of antibody responses in immunogenicity studies.
Main Methods:
- A high-throughput 96-well plate-based method using modified column chromatography.
- Utilized isotype-specific reagents for isolating swine IgG and IgA antibody isotypes.
- Collected various fluids (serum, milk, colostrum) from immunized pigs at multiple time points.
Main Results:
- Achieved rapid and consistent yield of specific IgG and IgA antibodies.
- Demonstrated comparable antibody isolation outcomes between single-column and 96-well plate systems.
- The 96-well plate system significantly reduced processing time compared to traditional methods.
Conclusions:
- The developed 96-well plate method offers a time-saving advantage for antibody isolation.
- This high-throughput approach is ideal for immunogenicity studies involving multiple animal samples and time points.
- The protocol effectively isolates swine IgG and IgA, facilitating sensitive antibody detection assays.
