Venom Peptide/Protein Purification

Venom is a complex mixture of inorganic and organic components, mainly proteins and peptides. Because the composition of the venom is complex and changeable due to various factors, it is very difficult to standardize the separation and purification method of biologically active molecules in the venom. Creative Proteomics aims to standardize a simple purification method to obtain high-purity peptides or proteins from venom, which is conducive to the study of the biochemical characteristics and functions of these molecules.

Introduction

The venom is composed of sodium, zinc, calcium and other ionic inorganic compounds, as well as organic components such as proteins and peptides which account for most of the venom. These toxins can act alone or in synergy, causing local or systemic tissue damage and various other toxic effects. However, the peptides or proteins in the venom are very complex and may contain hundreds of components. In order to separate and purify a specific protein from the venom, two or more chromatographic steps are usually required, which may include molecular repulsion, ion exchange, affinity, reverse phase, etc. And the choice of chromatographic type depends on the characteristics of each protein to be separated. Creative Proteomics provides customers with standardized separation and purification methods to obtain peptides or proteins in crude venom to facilitate future functional research.

Technical Process

We use chromatographic separation technology to obtain purified target peptides or proteins from the venom.

• Molecular exclusion chromatography. Purify the crude venom supernatant with the selected chromatographic column, select the target sample according to the protein profile of the eluted component in the gel, and freeze-dry it and submit it to the next chromatographic step.
• Ion exchange chromatography. Use a three-step elution, evaluate all the eluted components to screen out the chromatographic components with the target activity and lyophilize them.
• Ultrafiltration. After ultrafiltration, a sample of low molecular weight components such as salt is released, and then the concentration of the sample is determined and lyophilized.
• RP-HPLC. The purity level of the distillate after ultrafiltration was evaluated.

Result Delivery

Creative Proteomics can perform continuous chromatographic steps to separate and purify the venom peptide or protein component you are interested in, and provide satisfactory results, including but not limited to the following:

• Various chromatographic analysis charts and analysis results
• A target venom peptide or protein sample obtained by a continuous separation and purification method
• Purity and activity evaluation reports

Advantages

Our venom peptide or protein purification service usually uses 2 to 3 chromatographic steps, which has the advantages of molecular rejection and ion exchange. The one-stop service also provides you with follow-up identification services, including determination of molecular mass, amino acid sequence, and evaluation of functional activity.

Creative Proteomics can provide you with one-stop venom peptideomics and proteomics research services to help you conduct more comprehensive and in-depth venomomics research. If you want to know more about venom peptide and protein characterization services, please feel free to contact us.

Reference

• Danilo L, et al. Purification procedure for the isolation of a P-I metalloprotease and an acidic phospholipase A2 from Bothrops atrox snake venom. Journal of Venomous Animals and Toxins including Tropical Diseases, 2015.