High-throughput estimation of specific activities of enzyme/mutants in cell lysates through immunoturbidimetric assay of proteins

High Throughput Screening Technologies in Biomass Characterization

“…HTP screening methods are being increasingly applied to process development in biotechnology (Long et al, 2014;Back et al, 2016;Yang et al, 2017;Zutz et al, 2017). As new methodology is developed, HTP screening is increasingly employed to collect biological data that historically required extensive time and effort (Scheel and Lutke-Eversloh, 2013;Suzuki et al, 2015) and is widely used today in the development of fermentation process assays (Decker et al, 2003(Decker et al, , 2009Selig et al, 2010Selig et al, , 2011Studer et al, 2010;Suzuki et al, 2015;Yang et al, 2017). There have been rapid developments of HTP techniques in recent years in micro-scale culturing, online analysis and monitoring, and real-time control, which have enabled increased systems automation (Yang et al, 2017).…”

“…As new methodology is developed, HTP screening is increasingly employed to collect biological data that historically required extensive time and effort (Scheel and Lutke-Eversloh, 2013;Suzuki et al, 2015) and is widely used today in the development of fermentation process assays (Decker et al, 2003(Decker et al, , 2009Selig et al, 2010Selig et al, , 2011Studer et al, 2010;Suzuki et al, 2015;Yang et al, 2017). There have been rapid developments of HTP techniques in recent years in micro-scale culturing, online analysis and monitoring, and real-time control, which have enabled increased systems automation (Yang et al, 2017). A key technology in applying HTP to microbiological screening has been the miniaturization of bioreactors, making large experimental cultivation economical and practical (Back et al, 2016;Velez-Suberbie et al, 2017).…”

“…Recent HTP developments applied to biological research have been applied to developing more effective large-scale operations, greatly decreasing the time and expense compared to development at scale (Lattermann and Buchs, 2015) and it is likely that there is room for continued improvements (Long et al, 2014). As an example, microtiter plates (MTP) are simple, easy to shake, and inexpensive (Bharadwaj et al, 2011;Yang et al, 2017) and have been demonstrated to effectively replace shake flasks (Oguntimein et al, 2018). MTPs have been used to screen specific activities of enzyme variants using various techniques, such as protein quantitation by immunoturbidimetric (ITA) assays, (Yang et al, 2017), direct fluorescence resonance energy transfer for protease activity (Suzuki et al, 2015), cell free protein production (Casteleijn et al, 2013), fungal biosensor assay to detect estrogen activity (Zutz et al, 2017), protein purification and characterization for crystallographic studies (Kim et al, 2011), and enzyme-screening of ionic liquid pretreated lignocellulose (Bharadwaj et al, 2011).…”

“…As an example, microtiter plates (MTP) are simple, easy to shake, and inexpensive (Bharadwaj et al, 2011;Yang et al, 2017) and have been demonstrated to effectively replace shake flasks (Oguntimein et al, 2018). MTPs have been used to screen specific activities of enzyme variants using various techniques, such as protein quantitation by immunoturbidimetric (ITA) assays, (Yang et al, 2017), direct fluorescence resonance energy transfer for protease activity (Suzuki et al, 2015), cell free protein production (Casteleijn et al, 2013), fungal biosensor assay to detect estrogen activity (Zutz et al, 2017), protein purification and characterization for crystallographic studies (Kim et al, 2011), and enzyme-screening of ionic liquid pretreated lignocellulose (Bharadwaj et al, 2011). Despite these recent examples of MTP-based screening, few details are known regarding actual culture conditions inside the MTP, the technology to measure these details in real time in such numbers and small volumes remains lacking (Long et al, 2014).…”

Data for high-throughput estimation of specific activities of enzyme/mutants in cell lysates through immunoturbidimetric assay of proteins

“… Data in this article are associated with the research article “Highthroughput estimation of specific activities of enzyme/mutants in cell lysates through immunoturbidimetric assay of proteins” (Yang et al, 2017) [1] . This article provided data on how to develop an immunoturbidimetric assay (ITA) of enzyme/mutants as proteins in cell lysates in high-throughput (HTP) mode together with HTP assay of their activities to derive their specific activities in cell lysates for comparison, with Pseudomonas aeruginosa arylsulfatase (PAAS) and Bacillus fastidious uricase (BFU) plus their mutants as models.…”