HYBRIDOMA TECHNIQUES

Spring Quarter, 2007-2008

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Welcome to the homepage for this course. I hope that you will find this page to be useful as you as you prepare each week for Hybridoma Techniques lab. You can download and print each LAB procedure and any DISCUSSION NOTES from the links for each week's labs.

Each LAB procedure and set of DISCUSSION NOTES is also available as a PDF file. This format may be much easier to download and print. . Some links from PDF files do not work. ALL links from HTML files do work. The PDF files will require the Adobe Acrobat Reader to read PDF format. If you don't have a copy on your computer, click here for a free download. To go to links from PDF files, hold down the control key (Ctrl) while clicking on the link.

Hotbox
This box will contain announcements that need to be
passed along to students in Hybridoma Techniques

Lab Instructor
Kim Corbett
Offices: 08-1334
Phones: 475-6039
Email: kacsbi@rit.edu

You can send an email by simply clicking on my name or my email address above
to open an email dialogue box already addressed
OR
creating your own email message addressed to kacsbi@rit.edu


Suggested Readings
Monoclonal Antibodies
Monoclonal Antibodies (different site)
Monoclonal Antibody Production - The Basics
Uses for monoclonal antibodies
The Mouse in Science: Monoclonal Antibodies
Antibodies and therapy
Monoclonal Antibody Therapy
Some monoclonal antibodies that have been introduced into human medicine

Ethical Issues
Animal Welfare Act
(Animal Welfare Act - Summary PDF version)
AVMA Guidelines on Euthanasia (American Veterinary Medical Association, 2007)
Acceptable Methods for Euthanasia of Laboratory Animals at the University of Minnesota
Guide for the Care and Use of Laboratory Animals
The American Veterinary Association - Animal Welfare
Comparision of Ascites vs. Cell Culture for Production of Monoclonal Antibodies

Alternatives to Ascites Production of Monoclonal Antibodies

Tentative Grading

Resume Skills

Lab Safety Practices


Tentative Lab Schedule

Week #1 (March 10, 12)

LAB - Preparation of Lymphocyte Cell Suspension (PDF version)
LAB - Myeloma Cell Line Characteristics and Preparation (PDF version)

DISCUSSION NOTES - Introduction (PDF version)
DISCUSSION NOTES - Mouse Myeloma Cell Line (PDF version)


Week #2 (March 17, 19)

LAB - Fusion of Lymphocytes and Myeloma Cells (practice) (PDF version)

DISCUSSION NOTES - Fusion (PDF version)


Week #3 (March 24, 26)

LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)



Week #4 (March 31, April 2)

LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)

 

Week #5 (April 7, 9)

DISCUSSION NOTES - HAT Medium (PDF version)
DISCUSSION NOTES - 8-Azaguanine Resistance
(PDF version)
DISCUSSION NOTES - HT Medium (PDF version)

 

Week #6 (April 14, 16)

Quiz #1 (will cover Weeks 1-5)

LAB - Fusion of Lymphocytes and Myeloma Cells (PDF version)

 

Week #7 (April 21, 23)

LAB - Intraperitoneal Injection (PDF version) KLH, pristane, LAB - cells (PDF version)

DISCUSSION NOTES - Immunization of Mice (PDF version)
DISCUSSION NOTES - Intraperitoneal Injection of Mice (PDF version)

LAB - Intravenous Injection (PDF version) - NOT required, but worth trying

LAB - ELISA for the Detection of anti-KLH Antibodies (screening uncloned hybrids) (PDF version)

DISCUSSION NOTES - ELISA (PDF version)

 

Week #8 (April 28, 30)

LAB - Cloning by Limiting Dilution (PDF version)

DISCUSSION NOTES - Cloning by Limiting Dilution (PDF version)
DISCUSSION NOTES - Feeder Cells (PDF version)

 

Week #9 (May 5, 7)

LAB - ELISA for the Detection of anti-KLH Antibodies (screening cloned hybrids) (PDF version)

DISCUSSION NOTES - Pristane-Priming of Mice (PDF version)

 

Week #10 (May 12, 14)

Quiz #2 (will cover Weeks 6-9)

LAB - Transferring Cells to 24-well Plates and T-25 Flasks (PDF version)
LAB - Collection of ascites tumor cells from pristane-primed mice (demonstration only) (PDF version)

DISCUSSION NOTES - Expansion of cell culture volume (PDF version)