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Methods in Enzymology 2009This article reviews the development of immobilized-metal affinity chromatography (IMAC) and describes its most important applications. We provide an overview on the use... (Review)
Review
This article reviews the development of immobilized-metal affinity chromatography (IMAC) and describes its most important applications. We provide an overview on the use of IMAC in protein fractionation and proteomics, in protein immobilization and detection, and on some special applications such as purification of immunoglobulins and the Chelex method. The most relevant application-purification of histidine-tagged recombinant proteins-will be reviewed in greater detail with focus of state-of-the-art materials, methods, and protocols, and the limitations of IMAC and recent advances to improve the technology and the methods will be described.
Topics: Animals; Automation; Chromatography, Affinity; Equipment Reuse; Humans; Metals; Models, Biological; Protein Binding; Proteins
PubMed: 19892187
DOI: 10.1016/S0076-6879(09)63027-5 -
Methods (San Diego, Calif.) Mar 2017Antibodies continue to be extremely utilized entities in myriad applications including basic research, imaging, targeted delivery, chromatography, diagnostics, and... (Review)
Review
Antibodies continue to be extremely utilized entities in myriad applications including basic research, imaging, targeted delivery, chromatography, diagnostics, and therapeutics. At production stage, antibodies are generally present in complex matrices and most of their intended applications necessitate purification. Antibody purification has always been a major bottleneck in downstream processing of antibodies, due to the need of high quality products and associated high costs. Over the years, extensive research has focused on finding better purification methodologies to overcome this holdup. Among a plethora of different techniques, affinity chromatography is one of the most selective, rapid and easy method for antibody purification. This review aims to provide a detailed overview on affinity chromatography and the components involved in purification. An array of support matrices along with various classes of affinity ligands detailing their underlying working principles, together with the advantages and limitations of each system in purifying different types of antibodies, accompanying recent developments and important practical methodological considerations to optimize purification procedure are discussed.
Topics: Animals; Antibodies; Chromatography, Affinity; Humans; Hydrophobic and Hydrophilic Interactions; Ligands; Protein Binding; Recombinant Proteins; Static Electricity
PubMed: 28012937
DOI: 10.1016/j.ymeth.2016.12.010 -
Journal of Chromatography. B,... Nov 2020The field of affinity chromatography, which employs a biologically-related agent as the stationary phase, has seen significant growth since the modern era of this method... (Review)
Review
The field of affinity chromatography, which employs a biologically-related agent as the stationary phase, has seen significant growth since the modern era of this method began in 1968. This review examines the major developments and trends that have occurred in this technique over the past five decades. The basic principles and history of this area are first discussed. This is followed by an overview of the various supports, immobilization strategies, and types of binding agents that have been used in this field. The general types of applications and fields of use that have appeared for affinity chromatography are also considered. A survey of the literature is used to identify major trends in these topics and important areas of use for affinity chromatography in the separation, analysis, or characterization of chemicals and biochemicals.
Topics: Biochemistry; Biomedical Research; Biotechnology; Chromatography, Affinity; History, 20th Century; History, 21st Century; Humans
PubMed: 32871378
DOI: 10.1016/j.jchromb.2020.122332 -
Vaccine Aug 2019Affinity chromatography is among the most powerful separation techniques, achieving the finest separation with high yields even in the most challenging feed streams.... (Review)
Review
Affinity chromatography is among the most powerful separation techniques, achieving the finest separation with high yields even in the most challenging feed streams. Incorporating affinity chromatography in vaccine purification has long been attempted by researchers to improve unit yield and purity with the secondary goal of reducing the number of downstream process operations. Despite the success in laboratory-scale proof of concept, implementation of this technique in pilot or cGMP manufacturing has rarely been realised due to technical and economic challenges in design and manufacturing of ideal ligands as well as availability of high-productivity chromatography media. This paper reviews evolving technologies in engineered ligands and chromatography media that are encouraging companies to re-visit the possible use of affinity chromatography in larger scale vaccine purification. It is postulated that commercial-scale implementation of high throughput single-use affinity chromatography can significantly simplify process architecture, improve productivity and flexibility, and reduce cost of goods.
Topics: Automation; Bioreactors; Chromatography, Affinity; Ligands; Vaccines
PubMed: 29627235
DOI: 10.1016/j.vaccine.2018.02.090 -
Methods in Molecular Biology (Clifton,... 2015Affinity chromatography is one of the most selective and versatile forms of liquid chromatography for the separation or analysis of chemicals in complex mixtures. This... (Review)
Review
Affinity chromatography is one of the most selective and versatile forms of liquid chromatography for the separation or analysis of chemicals in complex mixtures. This method makes use of a biologically related agent as the stationary phase, which provides an affinity column with the ability to bind selectively and reversibly to a given target in a sample. This review examines the early work in this method and various developments that have lead to the current status of this technique. The general principles of affinity chromatography are briefly described as part of this discussion. Past and recent efforts in the generation of new binding agents, supports, and immobilization methods for this method are considered. Various applications of affinity chromatography are also summarized, as well as the influence this field has played in the creation of other affinity-based separation or analysis methods.
Topics: Chromatography, Affinity; History, 20th Century; History, 21st Century; Ligands
PubMed: 25749941
DOI: 10.1007/978-1-4939-2447-9_1 -
Biochimie Feb 2018The present review deals with the place of single chain oligonucleotide ligands (aptamers) in affinity chromatography applied to proteins. Aptamers are not the only... (Review)
Review
The present review deals with the place of single chain oligonucleotide ligands (aptamers) in affinity chromatography applied to proteins. Aptamers are not the only affinity ligands available but they represent an emerging and highly promising route that advantageously competes with antibodies in immunopurification processes. A historical background of affinity chromatography from the beginning of the discipline to the most recent outcomes is first presented. Then the focus is centered on aptamers which represent the last step so far to the long quest for affinity ligands associating very high specificity, availability and strong stability against most harsh cleaning agents required in chromatography. Then technologies of ligand selection from large libraries followed by the most appropriate chemical grafting approaches are described and supported by a number of bibliographic references. Experimental results assembled from relevant published paper are reported; they are selected by their practical applicability and potential use at large scale. The review concludes with specific remarks and future developments that are expected in the near future to turn this technology into a large acceptance for preparative applications.
Topics: Aptamers, Nucleotide; Chromatography, Affinity; Proteins
PubMed: 29054800
DOI: 10.1016/j.biochi.2017.10.008 -
Advances in Biochemical... 2020Aptamers are oligonucleotide molecules able to recognize very specifically proteins. Among the possible applications, aptamers have been used for affinity chromatography... (Review)
Review
Aptamers are oligonucleotide molecules able to recognize very specifically proteins. Among the possible applications, aptamers have been used for affinity chromatography with effective results and advantages over most advanced protein separation technologies. This chapter first discusses the context of the affinity chromatography with aptamer ligands. With the adaptation of SELEX, the chemical modifications of aptamers to comply with the covalent coupling and the separation process are then extensively presented. A focus is then made about the most important applications for protein separation with real-life examples and the comparison with immunoaffinity chromatography. In spite of well-advanced demonstrations and the extraordinary potential developments, a significant optimization work is still due to deserve large-scale applications with all necessary validations. Graphical Abstract Aptamer-protein complexes by X-ray crystallography.
Topics: Aptamers, Nucleotide; Chromatography, Affinity; Ligands; Proteins; SELEX Aptamer Technique
PubMed: 31485702
DOI: 10.1007/10_2019_106 -
Methods in Enzymology 1984
Review
Topics: Animals; Chromatography, Affinity; Cyanogen Bromide; Enzymes; Indicators and Reagents; Ligands; Resins, Synthetic
PubMed: 6371446
DOI: 10.1016/s0076-6879(84)04082-9 -
Expert Opinion on Drug Discovery Nov 2019: Fragment screening is a successful approach to accelerate drug discovery. Since it handles fragments with weak affinity, sensitive analytical tools are strongly... (Review)
Review
: Fragment screening is a successful approach to accelerate drug discovery. Since it handles fragments with weak affinity, sensitive analytical tools are strongly demanded. : After a short description of the available techniques employed in fragment screening, this review focuses on Weak Affinity Chromatography (WAC). Details include the origins of affinity chromatography and its evolution to WAC and the basic principles for affinity measurements. Preparation of columns for the immobilization of soluble target proteins and the development of lipodiscs for the incorporation of membrane proteins are described. The authors also discuss the advantages and limitations of WAC and compare it with other established techniques. : Although WAC is a suitable technique for fragment screening, with increased throughput, in particular, if coupled to mass spectrometry, it has not yet found the widespread application it deserves. A limiting factor may be the necessity for in house column preparation. In the future, the potential commercialization of columns with some important soluble protein targets would facilitate the application of WAC. The immobilization of membrane proteins on lipodisks is an area for further investigation, as is the comparison of WAC with other established methods since relevant information is still at a low level.
Topics: Chromatography, Affinity; Drug Discovery; Humans; Mass Spectrometry; Small Molecule Libraries
PubMed: 31380703
DOI: 10.1080/17460441.2019.1648425 -
Current Protocols in Protein Science Sep 2013Addition of an affinity tag is a useful method for differentiating recombinant proteins expressed in bacterial and eukaryotic expression systems from the background of... (Review)
Review
Addition of an affinity tag is a useful method for differentiating recombinant proteins expressed in bacterial and eukaryotic expression systems from the background of total cellular proteins, as well as for detecting protein-protein interactions. This overview describes the historical basis for the development of affinity tags, affinity tags that are commonly used today, how to choose an appropriate affinity tag for a particular purpose, and several recently developed affinity tag technologies that may prove useful in the near future.
Topics: Chromatography, Affinity; Molecular Probe Techniques; Recombinant Proteins
PubMed: 24510596
DOI: 10.1002/0471140864.ps0909s73