Biotechnology Techniques (biosc 275)
Biotechnology Techniques (Biosc 275)
Spring Quarter 2005
Instructor:
John
Wiseley
Lecture:
9:30-12:00 or 5:00-7:30 MW
Office:
2813
Room
2930
Telephone:
(206) 546-4588
Office Hours: 12:00-1:00, 4:00-5:00 MW
e-mail*:
jwiseley@shoreline.edu
* e-mail is the best way to contact me
Required Materials
Text: Fundamentals of Biochemistry (2002) by Voet, Voet & Pratt
Lab Guide: Experiments in Biochemistry (2000) by Farrell & Ranallo
Lab Notebook – Many types are possible, but must have bound pages
Prerequisites
BIOSC 201 Principles of Biology
BIOSC 250 Molecular Biology
CHEM 220 Organic Chemistry (strongly recommended)
Course Format
Biotechnology Techniques is a lab-centered course that will focus on current techniques
in protein purification and analysis. This will include cation-anion exchange
chromatography, selective binding using hydrophobic interactions, ultrafiltration,
isoelectric focusing and high performance liquid chromatography (HPLC). There will
also be exposure to good manufacturing practices (GMP), quality control procedures
(QC), biological potency assays, radioisotope use and handling, FDA regulations and
clinical trials.
In general, the first 30-45 minutes of class time will be devoted to lecture. Required
reading is indicated in the schedule. Additional reading materials will be provided as
needed.
BIOSC 275 – COURSE SCHEDULE – SPRING 2005
Revision may be needed during the quarter.
Week Date
Lecture Topic and Lab
Reading
1 Mon
Lecture: Acids, Bases and Buffers
Farrell & Ranallo (F&R)
3/28
Ch. 2 p. 37-50
Lab: Preparation of Buffers p. 51
Voit &Voit (V&V)
p. 31-37
Wed
Lecture: Spectrophotometry - Beer’s Law
F&R Ch. 3 p. 59-69
3/30
Protein Structure
V&V p. 77-86
Lab: Separation/Identification
of Amino Acids by TLC.
http://www.chm.davidson.e
p.126 part A only
du/ChemistryApplets/spectr
ophotometry/
2 Mon
Lecture: Enzyme Kinetics
F&R Ch. 8 p. 179-188
4/4
Lab: Determination of protein
V&V p. 281-287, 322-347
concentration p. 70
&
Kinetics of LDH – p. 199
Wed
Quiz #1: Amino Acids, Buffers, Specs
http://cti.itc.virginia.edu/~c
4/6
Lecture: Enzyme Kinetics (cont.)
mg/Demo/scriptFrame.html
– Michaelis-Menten,
Lab: Kinetics of LDH (cont.)
Lineweaver-Burke, and
Reversible Inhibition
3 Mon
Lecture: Enzyme Purification –
F&R Ch. 4 p. 83-94
4/11
Ammonium Sulfate Precipitation
V&V p. 93-104
Lab: A.S. precipitation of LDH
Comprehensive version p.103
Wed
Lecture: Enzyme Purification – Affinity
F&R Ch. 6 p. 143-147
4/13
Chromatography
Lab: Identification & concentration
of LDH fractions
4 Mon
Quiz #2 Enzyme Kinetics
F&R Ch. 7 p. 157-164
4/18
Lecture: Enzyme Purification – Affinity
Chromatography
Affinity Chromatography p. 148
Lab: Chromatography of LDH sample
Wed
Lecture: Gel Filtration Chromatography
F&R Ch. 7 p. 157-164
4/20*
Lab: Gel Filtration of LDH sample p. 169
5 Mon
Lab: Identification & concentration
4/25
of LDH fractions
Wed
Lecture: Electrophoretic Techniques
F&R Ch. 9 p. 207-217
4/27
Lab: SDS PAGE p. 229
V&V p. 104-121
Part 1 - setting up the gels
6 Mon
Quiz #3 Protein Purification
F&R Ch. 9 (cont)
5/2
Lecture: Electrophoretic Techniques
Lab: Electrophoresis of LDH fractions
Part 2 – running the gels
Wed
High Performance Liquid Chromatography
F&R Ch. 5 p. 111-124
5/4
(HPLC) - Ion Exchange Chromatography
Handouts
Lab: Introduction to hardware and
software
7
Mon
Lecture: HPLC continued
V&V p. 124-157
5/9**
(completion of this
Lab: Identification of isoenzyme
reading is primarily
Fractions - p. 133 (modified protocol) important for the final)
Wed
5/11
Lab: Identification of isoenzyme
Fractions - part 2 data analysis
8 Mon
Lecture: Two-dimensional Electrophoresis
Handouts
5/16
Part 1
Lab: 1st Dimension - Isoelectric
Focusing
Wed
Lecture: Two-dimensional Electrophoresis
5/18
Part 2
Lab: 2nd Dimension - SDS Page
Electrophoresis
9 Mon
Lecture: Introduction to virtual protein
V&V p. 288-319
5/23
purification.
Guest Speaker – Quality Control & Good
Manufacturing Practices
Wed
Quiz #4 Electrophoresis of Proteins
http://www.tlsu.leeds.ac.u
5/25
k/courses/bioc2060/protei
Lab: Complete All lab work
nlab102/proteinlab.html
10 Mon
MEMORIAL DAY – NO CLASS
5/30
Wed
Presentation and Discussion of
6/1
Virtual Protein Purification
LAB NOTEBOOKS DUE
11 Mon
Review for Final
6/6
Wed
Final Exam –
6/8
During Scheduled Class Time
*April 21st is the last day for the P/NC option
**May 9th is the last day to withdraw and receive a W grade
