Labrou N, Clonis Y D
Department of Agricultural Biology and Biotechnology, Agricultural University of Athens, Greece.
J Biotechnol. 1994 Aug 15;36(2):95-119. doi: 10.1016/0168-1656(94)90047-7.
The quality criteria imposed on several biochemicals are stringent, thus, high-separation purification technology is important to downstream processing. Affinity-based purification technologies are regarded as the finest available, and each one differs in its purifying ability, economy, processing speed and capacity. The most widely used affinity technology is classical affinity chromatography, however, other chromatography-based approaches have also been developed, for example, perfusion affinity chromatography, hyperdiffusion affinity chromatography, high-performance affinity chromatography, centrifugal affinity chromatography, affinity repulsion chromatography, heterobifunctional ligand affinity chromatography and the various chromatographic applications of 'affinity tails'. On the other hand, non-chromatographic affinity technologies aim at high throughput and seek to circumvent problems associated with diffusion limitations experienced with most chromatographic packings. Continuous affinity recycle extraction, aqueous two-phase affinity partitioning, membrane affinity filtration, affinity cross-flow ultrafiltration, reversible soluble affinity polymer separation and affinity precipitation are all non-chromatographic technologies. Several types of affinity ligands are used to different extents; antibodies and their fragments, receptors and their binding substances, avidin/biotin systems, textile and biomimetic dyes, (oligo)peptides, antisense peptides, chelated metal cations, lectins and phenylboronates, protein A and G, calmodulin, DNA, sequence-specific DNA, (oligo)nucleotides and heparin. Likewise, there are several support types developed and used; natural, synthetic, inorganic and composite materials.
对几种生物化学物质的质量标准要求严格，因此，高分离纯化技术对下游加工很重要。基于亲和作用的纯化技术被认为是现有的最佳技术，每种技术在纯化能力、经济性、处理速度和容量方面都有所不同。应用最广泛的亲和技术是经典亲和色谱法，不过，也开发了其他基于色谱的方法，例如灌注亲和色谱法、超扩散亲和色谱法、高效亲和色谱法、离心亲和色谱法、亲和排斥色谱法、异双功能配体亲和色谱法以及“亲和尾”的各种色谱应用。另一方面，非色谱亲和技术旨在实现高通量，并试图规避与大多数色谱填料所经历的扩散限制相关的问题。连续亲和循环萃取、水相双相亲和分配、膜亲和过滤、亲和错流超滤、可逆可溶性亲和聚合物分离和亲和沉淀均为非色谱技术。几种类型的亲和配体的使用程度各不相同；抗体及其片段、受体及其结合物质、抗生物素蛋白/生物素系统、纺织染料和仿生染料、（寡）肽、反义肽、螯合金属阳离子、凝集素和苯硼酸盐、蛋白A和G、钙调蛋白、DNA、序列特异性DNA、（寡）核苷酸和肝素。同样，也开发并使用了几种载体类型；天然材料、合成材料、无机材料和复合材料。
