Analysis of refolded antibody fragments by Differential Scanning Fluorimetry(DSF) (CAT#: STEM-MB-0796-WXH)

Name: Analysis of refolded antibody fragments by Differential Scanning Fluorimetry(DSF)
SKU: STEM-MB-0796-WXH
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Introduction

Fragment antibodies, as the name suggests, are fragments of the binding end of a monoclonal antibody (mAb) and can be a small but mighty tool to fight disease. While mAbs range from around 150kDa in size, fragment antibodies as small as 3kDa have been used in various applications. Refolding is one of the production technologies for pharmaceutical grade antibody fragments.
Differential scanning fluorimetry can be potentially employed as a qualitative measure of functionality, to evaluate refolded proteins, with applicability for antibody engineering in passive immunization.

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Principle Applications Procedure Materials

Principle

Differential Scanning Fluorimetry measures protein thermal unfolding by monitoring changes in fluorescence emission of a sample upon heating. This allows the determination of protein thermostability and complex formation even with weakly binding ligands by thermal shift assay. Differential Scanning Fluorimetry is therefore ideally suited for screening of optimum buffer conditions like pH, buffer composition and ionic strength. The technique is applicable to any biological sample, from soluble proteins to integral membrane proteins.

Applications

To identify low-molecular-weight ligands that bind and stabilize purified proteins.
To measure the denaturation and unfolding of proteins.

Procedure

1. Preparation of compound solutions
2. Preparation of buffer/additive screen plates
3. Preparation of compound storage plates
4. Equipment preparation
5. Sample preparation
50. Performing the scan