The Way We Take Care of Antibody Purificationby Creative Diagnostics Manufacturer & Supplier
The reason for us to purify antibody is to get usable antibody according to the intended applications, especially for high level of purification. A significant advantage for the purification of antibodies or antibody fragments, even from different sources, is to obtain a great deal of information available for the properties of the interested molecule and the major contaminants.
In the market, there are a lot of strategies can be reached, but either they are similar or variously different. In order to occupy a space, I guess each has to make best efforts. At Creative Diagnostics, as far as I know, the company will make following procedures to make sure ideal antibody purification outcomes:
1. Sample preparation
The reason to prepare samples before purification is to remove specific impurities, such as lipoproteins or phenol red, from the source material; or gross impurities, such as low molecular weight contaminants, bulk protein, from the source material and unwanted small molecules; or buffer exchange and desalting to make sure the correct buffer conditions (for example, pH and salt concentration). Usually in laboratory condition, the buffer exchange and desalting can be skipped if reasonable procedures were conducted after filtration and centrifugation, such as affinity chromatography.
2. Rigorous purification by affinity chromatography
Due to high selectivity and subsequently high resolution, and high capacity for the target proteins, affinity chromatography is usually applied in line with the immunospecific interaction. For polyclonal antibodies (serum samples), for example, the antigen-specific affinity purification is necessary to prevent co-purification of nonspecific immunoglobulins. While newly emerged changes, such as recently advanced technologies in the production and purification techniques, genetically engineered antibodies and antibody fragments as one of such examples, have brought log of opportunities for the antibody purification. By so, tags can be introduced into target molecules without affinity media.
In the process of using immunoprecipitation, we need large quantity of subjective experience and objective expertise to obtain satisfactory results.
4. Mutiple statigies to obtain purification
As previously discussed in part two, a rigorous but rapid purification step is generally enough to get the ideal level of purity and quantity of product required for research purposes by using affinity chromatography. Through this, antibodies or antibody fragments can be adequately purified for next use, with following step (such as desalting, buffer exchange and high resolution gel filtration) to get out of unwanted impurities. But in case, if an affinity chromatography is not available or if a higher degree of purity is required, Creative Diagnostics will conduct alternative techniques (combined effectively).
5. Apply further purification after intial procedures to remove specific contaminants.
6. Both small and large scale purification is attainable.
About Creative Diagnostics
Creative Diagnostics staff scientists have extensive antibody purification experience. It has developed robust proprietary purification procedures for antibodies from serum, ascites fluid, yolks and culture supernatants for use in early discovery research, preclinical trials and in vitro medical devices. All purified antibodies can be analyzed by electrophoresis, ELISA, western blot and HPLC to determine purity and integrity.
Created on Dec 31st 1969 19:00. Viewed 0 times.