Основы белковой хроматографии
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Applications that requires biochromatography, a few examples
Mass Spectrometry Analysis of partners/Complexes/petide structure
–Requires less proteins, but start from large samples
–No Tags, work in the endogenous species
–LC-MS/MS or MALDI–TOF in downstream
Step 1: Cell |
Step 2: |
Step 3: Perform |
Culture (or |
Chromatographically MS analysis |
|
tissue) |
purify crude lysates |
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and verify by SDS- |
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PAGE |
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Life Science Group I Chromatography
Applications that requires biochromatography, a few examples
Monoclonal Antibodies (mAbs) Purification
–The goal is to purify large amount of Antibodies
–Multiple Monoclonal cell lines are cultured, to find the best expressing cell line
–Typical labs are Pharma or Antibody producing Biotechs
Step 1: |
Step 2: |
Step 3: Perform |
|
Hybridoma Cell Chromatographically |
Antigen- |
||
Culture |
purify crude lysates |
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Antibody |
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and verify by SDS- |
|
interaction |
|
PAGE |
|
analysis |
Life Science Group I Chromatography
Applications that requires biochromatography, a few examples
Cellular extract Purification for Biotherapeutics Discovery
Goal: Identify new active molecules from natural products
Step 1 |
Step 2 |
Step 3 |
Extraction from |
Fractions are |
Fractions are |
biologically active |
separated. Typical |
tested for |
plant or animal |
methods: Ion |
biological activity. |
tissues. |
exchange, Size |
Sub-fractions can |
|
exclusion. |
be isolated further. |
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Life Science Group I Chromatography
Thank You Very much for atention! Questions?
Life Science Group I Chromatography