Stability of the lyophilized F(ab’)2 fragments of horse tetanus antibodies isolated by affinity chromatography.

The research paper is about a study on the stability of lyophilized F(ab’)2 fragments of horse tetanus antibodies, which found that these fragments remain stable even after being subjected to a freeze-drying process.

Study Overview

This study involved the examination of the F(ab’)2 fragments of horse tetanus antibodies and how stable they were when subjected to lyophilization, a freeze-drying process. Several methods and substances were used during this study, including peptic digestion, affinity chromatography, ultrafiltration, and the use of glycine and human serum albumin as stabilizing agents.

• The researchers first removed the F(ab’)2 fragments from horse hyperimmune sera through peptic digestion.
• These fragments were then passed through a column of tetanus toxoid coupled with Sepharose 4B.
• The F(ab’)2 fragments were then extracted using a solution of 5 mM HCl in 150 mM NaCl.
• This was followed by the concentration of the eluates via ultrafiltration, after which they were lyophilized.
• Glycine and human serum albumin were utilized as stabilizing agents during the study.

Findings

The researchers discovered that both the polyacrylamide gel electrophoretic mobility and the molecular weight of the F(ab’)2 fragments remained the same after they had been lyophilized. Furthermore, the preparations, when stored two months at 56 degrees Celsius, showed only a slight decrease in antitetanus activity. Observations from ORD measurements and spectrophotometric determinations in the pH range 2.1-5.0 indicated that the structure of the F(ab’)2 fragment is highly stable in an acid medium.

• The F(ab’)2 fragments maintained their polyacrylamide gel electrophoretic mobility and molecular weight after being subjected to lyophilization.
• After two months of storage at 56 degrees Celsius, there was only a minor decrease in antitetanus activity in the freeze-dried preparations.
• ORD measurements and spectrophotometric determinations over a pH range of 2.1-5.0 showed the F(ab’)2 fragment structure to have high stability in an acidic environment.

Significance

The findings of this research are significant as they show that the freeze-drying process, or lyophilization, can be used on F(ab’)2 fragments of horse tetanus antibodies without compromising their stability. It demonstrates the viability of this method for long-term storage and transportation of these fragments, which have important applications in immunity and disease treatment.