Publication Date

Summer 2011

Document Type

Project Summary

Degree Name

Master of Science

Department

Analytical Chemistry

First Advisor

Walter Henne, Jr., Ph.D.

Second Advisor

Joseph B. Addison, Ph.D.

Third Advisor

Aheda Saber, Ph.D.

Abstract

The main objective of this project was to develop and validate methods for detection and analysis of proteins using a newly introduced microdrop plate reader equipped with a 16 well microdrop reader (2 uL sample volume). We used commercially available BSA, egg albumin, trypsinogen, Pepsin, β-lactoblobulin and lysozyme as out test moieties. Protein drop size, time, BCA reagent quantity, was assessed at various levels using a microdrop micro-BCA protocol (Pierce Chemical Company) with the appropriate controls (PBS blank) the method validated using the appropriate statistical and method parameters (i.e %RSD, and regression analysis of the standard curves). The micro method was used to determine the concentration of 5 known proteins and the method compared to the standard 96 well BCA plate method. The overarching strategy in our group is to develop, simple robust assays that use minimal sample amounts and reagents given the high cost and limited availability of many proteins and reagents (not to mention limiting waste disposal). Moreover, the technique will aid researches in quickly identifying and analyzing proteins during expression, analysis, and isolation.

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