The purpose of this research was to develop a laboratory procedure using an inexpensive surface plasmon resonance instrument for undergraduate use that investigates the binding of antigens and antibodies, specifically α-Lactalbumin and anti-α-Lactalbumin. In order for antigen-antibody interactions to be studied by SPR, antibodies must first be attached to a gold-coated glass slide. 2-Mercaptoethylamine (MEA) was used to break the disulfide bonds of the antibodies so they could be attached to the gold-plated slide by exposed thiols on the F<sub>c</sub> tail of the molecule. The fragmented antibodies stuck to the gold surface of the slide and produced a consistent change in the SPR angle. As the antigen solution was slowly pumped through the sample cell and over the antibodies, a further change in SPR angle was expected. However, the observed change was inconsistent. In some trials the SPR angle increased while in others it decreased. Our instrument was not programmed to generate a graph of the response as a function of time—called a sensorgram. Thus, there was no way to determine whether or not the α-Lactalbumin bound to the antibodies, and an analysis of the antigen-binding properties could not be completed. The inexpensive SPR instrument could be used by undergraduates to gain familiarity with the SPR technique, but did not prove to be as useful as hoped for purposes of studying antigen-antibody dynamics.

Use of an inexpensive surface plasmon resonance instrument to determine the binding of α-lactalbumin to fragmented antibodies

2019 Spring National Meeting

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