Purification of antibodies by affinity chromatography

Antibadiler tedavi amaçlı uygulamalarda yoğun olarak kullanılan bir protein türüdür. Antibadiler değişik saflaştırma tekniklerinin bir arada kullanımı ile saflaştırılırlar. En yaygın kullanılan saflaştırma tekniği immunoglobulin G’nin (IgG) afinite ligand ile seçici etkileşimine dayanan afinite kromatografi tekniğidir. Afinite kromatografisinin kolay, hızlı ve hedef molekülü seçici olarak yakalama gibi birçok avantajları söz konusudur. Bu derleme makalede son yıllarda antibadi saflaştırma amacıyla kullanılan farklı afinite teknikleri ve elde edilen önemli sonuçları verilmiştir. Bu kapsamda temel ayırma teknikleri olarak; protein afinite kromatografisi, boyaligand afinite kromatografisi, immobilize metal-şelat afinite kromatografisi, karışık-mod afinite kromatografisi, tiyofilik kromatografi, manyetik olarak kararlaştırılmış akışkan yataklar ve sulu iki fazlı sistemler ve önemli sonuçları tartışılmıştır.

Afinite kromatografisi ile Antibadi saflaştırılması

Antibodies are becoming a dominant protein class for various human therapeutical applications and they represent the largest number of molecules in clinical trials today. Antibodies are purified using a combination of various purification techniques. The most popular technique is affinity chromatography based on specific interaction of immunoglobulin G (IgG) with affinity ligand. Affinity chromatography has several advantages since it is an easy, fast and selective procedure for capturing the target antibody. This review aims at describing different affinity techniques for purification of antibodies and reports the contributions of scientists working in this field and their important results. The main issues discussed in this review are: protein A affinity chromatography, histidine-ligand affinity chromatography, immobilized metal-chelate affinity chromatography, dye-ligand affinity chromatography, biomimetic affinity chromatography, negative chromatography, mixed-mode affinity chromatography, thiophilic chromatography, magnetically stabilized fluidized beds and aqueous two phase systems.

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