UNIVERSITY OF ZIMBABWE

2018 Nov/Dec Examinations

Science
Faculty:...................................................................................................................................
Biochemistry
Department:............................................................................................................................
HBC201: Practical Techniques in Biochemistry 2
Paper code and Title:............................................................................................................
2 hours
Duration:.................................................................................................................................
Mr F. Munyonho
Examiner:................................................................................................................................

Authorized Materials:

INSTRUCTIONS:

1. This paper contains 6 Questions

2. Answer 4 questions
3. Start each question on a new page
4. This question paper comprises 2 printed pages

NB: DO NOT TURN OVER THE QUESTION PAPER OR COMMENCE WRITING UNTIL
INSTRUCTED TO DO SO.
HBC201 PAGE ONE

1. Discuss how a mixture of non-volatile compounds maybe separated by gas

chromatography. [25 marks]

2.

(a) Write notes on the mobile phases used for HPLC analysis. [12 marks]

(b) Give examples of clinical applications of the Western blot technique. [5 marks]

(c) Briefly describe any FOUR factors that may affect the migration of ions during
electrophoresis. [8 marks]

3. With the aid of labeled diagrams, outline the steps in the following types of ELISA:

(a) sandwich [12 marks]

(b) competitive [13 marks]

4.
(a) Using an example, describe the basis of separation of biomolecules by
by ion exchange chromatography [10 marks]

(b) The table below shows the list of proteins in a sample, and their respective
molecular mass and pI values.

Protein Mr pI
A 12, 000 3
B 28, 000 8.4
C 42, 000 7.2
D 66, 000 9
E 111, 000 4.2

Assuming that the protein remains “native,” or non-denatured, explain the

order in which the proteins will elute, when separated using the following
techniques:

(i) Anion exchange chromatography, pH 9, using the fusing salt gradient to elute
[8 marks]
PLEASE TURN TO THE NEXT PAGE
HBC201 PAGE TWO

(ii) Cation exchange chromatography, pH 4.2, using salt gradient to elute

[7 marks]

5.
(a) Using concanavalin A as an example, describe how one would isolate a lectin
from a crude extract. [13 marks]

(b) Discuss the limitations of using dialysis in the purification of biomolecules.

[12 marks]

6.
(a) Write notes on the focusing effect, as used in the IEF electrophoresis. [5 marks]

(b) Briefly describe the following methods including their respective applications in
biotechnology:

(i) 2D gel electrophoresis;

(ii) SDS polyacrylamide gel electrophoresis (PAGE). [10 marks each]

END OF QUESTION PAPER