Apronex

1. Bacterial paste or cell pellet lysate preparation

Cells are disrupted mechanically (Sonication, French Press)  or with a detergent, centrifuged and the soluble and insoluble fractions are separated and collected.

2. Protein extraction from insoluble pellet/membranes

Proteins are extracted from insoluble material using customer supplied protocol, or protocol selected by Apronex We offer various extraction options to suite most target proteins. If needed, an extraction step can be developed by testing different extraction conditions on a small scale.

3. Refolding of solubilized inclusion bodies

The refolding conditions are tested.

4. Chromatography

Chromatography steps follow customer-supplied protocols, published protocols, standard protocols (for tagged/fused proteins only) or protocols developed by Apronex Our standard chromatography modes include gel filtration, ion exchange, tag affinity, immunosorbents, hydrophobic interaction. Dye affinity, nucleotide affinity, hydroxylapatite and other chromatography modes can be used if needed. Each run is performed on a dedicated column. Mostly we use resin filled columns from Bio-Rad and for large scale purifications special custom-made columns are prepared. Purification steps are monitored by light absorbance at 280 nm. Fractions are analyzed by SDS-PAGE/Protein stain or other assays can be performed when necessary.

5. Endotoxin removal

Endotoxin is removed from protein preparations.