Homework Help is Here – Start Your Trial Now!
Science
Biochemistry
If there are still contaminating proteins with your LDH fraction after going through 3 different purification schemes (affinity, ion exchange, and size exclusion columns), what would be their physical characteristics?

If there are still contaminating proteins with your LDH fraction after going through 3 different purification schemes (affinity, ion exchange, and size exclusion columns), what would be their physical characteristics?

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
See similar textbooks
icon
Related questions
Question
100%
**Title: Understanding Contaminant Proteins in LDH Fraction After Purification** **Question:** If there are still contaminating proteins in your LDH (Lactate Dehydrogenase) fraction after going through three different purification schemes (affinity, ion exchange, and size exclusion columns), what would be their physical characteristics? **Explanation:** After subjecting your LDH fraction to three purification methods, ideally, contaminants should have been removed. However, if they persist, they likely possess similar physical characteristics to LDH, making them difficult to separate. Below are explanations of each purification scheme and potential reasons for contamination. **1. Affinity Chromatography:** - **Principle:** Relies on the specific binding affinity between a protein and a particular ligand. - **Potential Issues:** Contaminants may share a similar affinity for the ligand used, resulting in co-elution with LDH. **2. Ion Exchange Chromatography:** - **Principle:** Separates proteins based on their net charge by using a charged resin. - **Potential Issues:** Contaminants may have similar charge properties to LDH under the chosen pH and ionic strength conditions, leading to incomplete separation. **3. Size Exclusion Chromatography (SEC):** - **Principle:** Separates proteins based on size as they pass through a column filled with porous beads. - **Potential Issues:** Proteins of similar molecular size may remain inseparable from LDH, as SEC does not distinguish between proteins of similar sizes or native conformations. **Conclusion:** Understanding the characteristics of contaminants is essential. They likely share specific binding, charge, or size properties with LDH, complicating their removal. Further purification strategies may involve adjusting pH, ionic conditions, or using more specific affinity tags.
Transcribed Image Text:**Title: Understanding Contaminant Proteins in LDH Fraction After Purification** **Question:** If there are still contaminating proteins in your LDH (Lactate Dehydrogenase) fraction after going through three different purification schemes (affinity, ion exchange, and size exclusion columns), what would be their physical characteristics? **Explanation:** After subjecting your LDH fraction to three purification methods, ideally, contaminants should have been removed. However, if they persist, they likely possess similar physical characteristics to LDH, making them difficult to separate. Below are explanations of each purification scheme and potential reasons for contamination. **1. Affinity Chromatography:** - **Principle:** Relies on the specific binding affinity between a protein and a particular ligand. - **Potential Issues:** Contaminants may share a similar affinity for the ligand used, resulting in co-elution with LDH. **2. Ion Exchange Chromatography:** - **Principle:** Separates proteins based on their net charge by using a charged resin. - **Potential Issues:** Contaminants may have similar charge properties to LDH under the chosen pH and ionic strength conditions, leading to incomplete separation. **3. Size Exclusion Chromatography (SEC):** - **Principle:** Separates proteins based on size as they pass through a column filled with porous beads. - **Potential Issues:** Proteins of similar molecular size may remain inseparable from LDH, as SEC does not distinguish between proteins of similar sizes or native conformations. **Conclusion:** Understanding the characteristics of contaminants is essential. They likely share specific binding, charge, or size properties with LDH, complicating their removal. Further purification strategies may involve adjusting pH, ionic conditions, or using more specific affinity tags.
Expert Solution
trending now

Trending now

This is a popular solution!

steps

Step by step

Solved in 2 steps

SEE SOLUTIONCheck out a sample Q&A here
Blurred answer
Similar questions
Recommended textbooks for you
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781319114671
Author:
Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:
W. H. Freeman
Lehninger Principles of Biochemistry
Lehninger Principles of Biochemistry
Biochemistry
ISBN:
9781464126116
Author:
David L. Nelson, Michael M. Cox
Publisher:
W. H. Freeman
Fundamentals of Biochemistry: Life at the Molecul…
Fundamentals of Biochemistry: Life at the Molecul…
Biochemistry
ISBN:
9781118918401
Author:
Donald Voet, Judith G. Voet, Charlotte W. Pratt
Publisher:
WILEY
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781305961135
Author:
Mary K. Campbell, Shawn O. Farrell, Owen M. McDougal
Publisher:
Cengage Learning
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781305577206
Author:
Reginald H. Garrett, Charles M. Grisham
Publisher:
Cengage Learning
Fundamentals of General, Organic, and Biological …
Fundamentals of General, Organic, and Biological …
Biochemistry
ISBN:
9780134015187
Author:
John E. McMurry, David S. Ballantine, Carl A. Hoeger, Virginia E. Peterson
Publisher:
PEARSON
SEE MORE TEXTBOOKS