Transcript see how is Zestern analysis different from current existing

Zestern analysis
A new method to address crossreactivity of antibodies
Cross-reactivity of antibodies
One antibody, one protein (antigen) is the basis of immunoblot
analysis
One antibody, two or more proteins (antigens) is the reality.
Majority of antibodies for research purpose actually recognize more than one
protein (Cross-reactivity)*.
*531 out of 22,000 antibodies are found without cross-reactivity
Schwenk et al. Mol. & Cell Proteomics 9:2497-2507,2010.
cross-reactivity of antibodies
reflected in Western blot analysis
Antibody w/o
Cross-reactivity
Antibody w/
Cross-reactivity
Traditional methods to address crossreactivity of antibodies
• Western blot analysis: Gel electrophoresis
The number of samples can be analyzed are
limited due to gel electrophoresis step
• Dot blot analysis
• ELISA
• Protein microarrays
Extensive screening to identify
antibodies without cross-reactivity
The number of suitable antibodies are limited.

Insufficient coverage

high developmental cost
Introducing Zestern analysis
Basic of Zestern analysis
Excessive antigen
Excessive partial antigen
Synthesized epitope peptide
Synthesized epitope repeat Peptide
Antibody
Antigen
Nitrocellulose
membrane
Epitope
How Zestern analysis achieves
specificity
Antibody with
cross-reactivity
a
a
A
b
B
B
B
A
a
A
Quantification
directly in solution
A
Antibody w/o
cross-reactivity
Antibody w/
cross-reactivity
Antigen A
B
Antigen B
a
Epitope a
b
Epitope b
b
a
b
a
A
Antibody without
cross-reactivity
b
B
Synthesized
epitope
Advantages over traditional
immunoblot analysis
• Eliminates gel electrophoresis step in Western
Blot analysis.
• Increases number of antibodies available for Dot
blot, ELISA and protein microarray analysis.
• Increases the coverage of proteins for Dot blot,
ELISA and protein microarray analyses.
• Serves as foundation for high throughput protein
analysis.
• Provides basis for automation of immunoblot
analysis.
Further Info.
• Zestern Biotechnique LLC owns US patent for Zestern
analysis.
• PCT application has been filed by Zestern
Biotechnique LLC.
• For further inquiry, contact:
• Dr. Jiandi Zhang
• [email protected]
• (001)-703-425-6666.