Leveraging protein purification strategies in proteomics

Roy, Ipsita; Mondal, Kalyani; Gupta, Munishwar N.

doi:10.1016/j.jchromb.2006.11.016

Cited by 34 publications

(29 citation statements)

References 185 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It has been recently reported that optimized ATPS can serve as a presorting stage in proteomic studies since it can be customized for selective extraction and/or partition of large fractions of proteins from crude extracts. These recent applications include 3D protein characterization when coupled with 2D electrophoresis [26,32]. Two-dimensional gel technology offers protein visualization for a variety of purposes, in addition to allowing empirical estimation of molecular weight, pI and expression levels [33].…”

Section: Atps and 2d Electrophoresis For 3d Characterization Of Proteinsmentioning

confidence: 99%

Extraction and Purification of Bioproducts and Nanoparticles using Aqueous Two‐Phase Systems Strategies

et al. 2008

View full text Add to dashboard Cite

Aqueous Two-Phase Systems (ATPS) is a primary recovery technique that has shown great potential for the efficient extraction and purification of high value biological compounds. The main advantages of this technique include scaling up feasibility, process integration capability and biocompatibility. In this review, the efficient use of ATPS for the extraction of proteins, genetic material, low molecular weight compounds, bioparticles, nanoparticles and cells is highlighted. The important role of ATPS in process integration, i.e., extractive conversion, extractive fermentation, cell disruption integrated with product recovery, and extractive purification, is discussed. A novel approach to protein molecular characterization combining ATPS and 2-dimension electrophoresis (2-DE) is introduced as a first step in the process development. Novel approaches for downstream processing using ATPS and dielectrophoresis are presented. Finally, trends concerning the application of ATPS strategies to address the future challenges of bioseparation are discussed.

show abstract

Section: Atps and 2d Electrophoresis For 3d Characterization Of Proteinsmentioning

confidence: 99%

Extraction and Purification of Bioproducts and Nanoparticles using Aqueous Two‐Phase Systems Strategies

et al. 2008

View full text Add to dashboard Cite

show abstract

“…Affinity chromatography, rather than being a 'polishing' step at the end, was promoted to the earlier steps of purification. In fact, upstream and downstream processing steps were integrated (Gupta, 2002;Przybycien et al, 2004;Mondal et al, 2006). (2) It was realized that enzymes could be used in non aqueous media (Carrea and Riva 2000; Gupta 2000;Patel 2000; Vulfson et al, 2001).…”

mentioning

confidence: 99%

“…Use of the affinity based separation strategies played an important role in this landmark innovation. Fusion tags (most familiar example is poly His tag) enable direct application of affinity chromatography called immobilized metal affinity chromatography (IMAC) (Porath, 1975;Mbrabet and Vijaylakshmi, 2002;Mondal and Gupta, 2006).While the trend is to use available commercial kits or commercially available affinity media; it helps to understand the design of the affinity media. In general, an affinity matrix is a bioconjugate of an affinity ligand which is attached to a polymeric matrix.…”

mentioning

confidence: 99%

“…The target protein recognizes the affinity ligand, binds it in a somewhat selective fashion and can be eluted often as a pure protein. Often, even the elution protocol can bring in considerable selectivity (Mondal et al, 2006). With poly His-tagged proteins, the metal releases a coordinated ligand (like, even the water molecule) and complexes with imidazole side chains of the histidine residues.…”

mentioning

confidence: 99%

“…A special mention must be made, to the use of smart polymers in affinity based separations. As most of the early work in this area has been reviewed periodically including an earlier review in this very journal (Roy and Gupta, 2002a), the discussion here will be limited to briefly mentioning recent work on simultaneous purification and refolding of proteins by using either affinity precipitation or macro-affinity facilitated three phase partitioning (MLFTPP) (Mondal et al, 2003;Mondal and Gupta, 2006;Mondal et al, 2006;Gautam et al, 2012).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Skills in biocatalysis are essential for establishing a sound biotechnological base in India

Gupta¹

2015

Proceedings of the Indian National Science Academy

View full text Add to dashboard Cite

Techniques developed by enzymologists and protein chemists have been indispensible in many areas of biotechnology. Bio-separation of proteins, biocatalyst engineering and medium engineering are important examples of this. Our understanding of enzyme specificity has helped us in gaining insight into catalytic promiscuity. This, along with the possibility of carrying out biocatalysis in organic solvents has widened the scope of using enzymes for organic synthesis.Reestablishing a strong base in enzymology is crucial towards creating a strong base for the field of biotechnology in India. Keywords IntroductionQuite a few aspects of biotechnology involve production and applications of enzymes. In the old days, most of these aspects were covered under industrial enzymology (Godfrey and Reichelt, 1983). Gradually, a more broad based term 'applied biocatalysis' came into vogue (Straathof and Adlercreutz, 2000) which was an interface between enzymology and biochemical engineering. This phase was characterized by few new developments: (1) As a result of the rDNA technology, our capability to produce mutant properties grew exponentially. There was a scramble to produce proteins at a lower cost. The stakes were no longer limited to low cost industrial grade enzymes. Costlier proteins used in molecular biology, diagnostics and biosensors and pharmaceutical proteins/enzymes could be produced by cloning and their properties tailored to suit the applications. The result was a paradigm shift in the downstream processing methods. Shorter and efficient protocols emerged. Affinity chromatography, rather than being a 'polishing' step at the end, was promoted to the earlier steps of purification. In fact, upstream and downstream processing steps were integrated (Gupta, 2002;Przybycien et al., 2004;Mondal et al., 2006). (2) It was realized that enzymes could be used in non aqueous media (Carrea and Riva 2000; Gupta 2000;Patel 2000; Vulfson et al., 2001). This opened up novel applications; especially in the synthesis of chiral compounds. Table 1 lists some of the milestones in the area of applied biocatalysis.It is remarkable how much enzymology contributed to all this.The present review attempts to emphasize the importance of the skills which are part of the toolbox Published Online on 10 December 2015 1114Munishwar Nath Gupta and Joyeeta Mukherjee et al., 1973; Mosbach 1976 Mosbach , 1980 Nilsson and Mosbach 1981; Mattiasson 1983;Mattiasson 1988) Berezin's group in Moscow was the third one (Berezin 1978; Mozhaev et al., 1987;Martinek and Mozhaev 1993). Current work is adequately discussed at many places ( Enzyme deactivation models Most people wrongly give half lives without analysing the kinetics of deactivation. Sadana's work needs to be known more widely. (Sadana 1991(Sadana , 1993 Enzymes as reverse micelles This is another approach in low water enzymology. (Luisi 1985; Larsson et al., 1987; Adlercreutz et al., 1988) Enzyme use in biosensors Immobilization allows enzymes to interface with transducers. (Yarapolov et al...

show abstract

Aqueous Two‐Phase Systems (ATPSs) for Extraction and Purification of Biomolecules and Bioparticles

Marques

Molino

Mazzola

et al. 2013

Encyclopedia of Industrial Biotechnology

View full text Add to dashboard Cite

The study of biomolecules and bioparticles has achieved great advances, thus increasing the productivity with upstream improvements. Nowadays, downstream processes are the bottleneck in the production of some biopharmaceuticals, a change from the previous years' scene. Current purification platforms will reach their physical limits at some point, creating the necessity for new approaches. This article reviews an alternative method for the extraction and purification of biomolecules/bioparticles denominated as aqueous two‐phase systems (ATPSs) . Biocompatibility and readiness to scale up are some of the characteristics of ATPSs and we also discuss its application in the biotechnology field.

show abstract

Leveraging protein purification strategies in proteomics

Cited by 34 publications

References 185 publications

Extraction and Purification of Bioproducts and Nanoparticles using Aqueous Two‐Phase Systems Strategies

Extraction and Purification of Bioproducts and Nanoparticles using Aqueous Two‐Phase Systems Strategies

Skills in biocatalysis are essential for establishing a sound biotechnological base in India

Aqueous Two‐Phase Systems (ATPSs) for Extraction and Purification of Biomolecules and Bioparticles

Contact Info

Product

Resources

About