INTRODUCTION

Proteins are important biomolecules. They have numerous functions and these are in accordance with
their specific structure and chemical composition. Basically, proteins contain subunits of polypeptides,
which contain sequences of amino acids. Proteins are often isolated by different techniques based on
different principles, like chemistry, solubility or even particle size. They are also hydrolyzed by acids,
bases and enzymes to be able to gain information on their composition. This is where the proteins are
cleaved into peptide and amino acid fragments. The isolated (isolate) and hydrolyzed(hydrolysate)
proteins are in turn, further analyzed quantitatively and qualitatively. The concentration is also determined
through methods of total protein analysis wherein the principle of the direct relationship of absorbance to
concentration is utilized. Analysis of the different forms of proteins are important especially for
pharamceutical industries or even nutrition companies which produce supplements and enhancing
substances.

The objectives of the experiment are to demonstrate commonly used protein isolation techniques and
explain the principles involved in each isolation technique, perform qualitative tests on the amino acids in
intact and hydrolyzed proteins and explain the principles involved in each test, execute acid, alkaline, and
enzymatic hydrolysis on the isolated proteins and enumerate the advantages and disadvantages of using
each type of hydrolysis, determine the amino acid components of proteins using thin-layer
chromatography and quantitatively determine the protein concentration of a given sample using
commonly used and microscale methods of total protein analysis.

Sources:

What are proteins and what do they do.(2017). National Library of Medicine. Retrieved from
https://ghr.nlm.nih.gov/primer/howgeneswork/protein

Protein Purification and Isolation.(n.d.). Bio-Rad. Retrieved from http://www.bio-rad.com/en-

us/applications-technologies/protein-purification-isolation