@article{FischbachLohBieretal.2017, author = {Fischbach, Jens and Loh, Qiuting and Bier, Frank Fabian and Lim, Theam Soon and Frohme, Marcus and Gl{\"o}kler, J{\"o}rn}, title = {Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method}, series = {Scientific Reports}, volume = {7}, journal = {Scientific Reports}, issn = {2045-2322}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-9248}, year = {2017}, abstract = {We identified Alizarin Red S and other well known fluorescent dyes useful for the online detection of pyrophosphate in enzymatic assays, including the loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. An iterative screening was used for a selected set of compounds to first secure enzyme compatibility, evaluate inorganic pyrophosphate sensitivity in the presence of manganese as quencher and optimize conditions for an online detection. Of the selected dyes, the inexpensive alizarin red S was found to selectively detect pyrophosphate under LAMP and PCR conditions and is superior with respect to its defined red-shifted spectrum, long shelf life and low toxicity. In addition, the newly identified properties may also be useful in other enzymatic assays which do not generate nucleic acids but are based on inorganic pyrophosphate. Finally, we propose that our screening method may provide a blueprint for rapid screening of compounds for detecting inorganic pyrophosphate.}, language = {en} } @article{DevulapallyBuergerMielkeetal.2018, author = {Devulapally, Praneeth Reddy and B{\"u}rger, J{\"o}rg and Mielke, Thorsten and Konthur, Zolt{\´a}n and Lehrach, Hans and Yaspo, Marie-Laure and Gl{\"o}kler, J{\"o}rn and Warnatz, Hans-J{\"o}rg}, title = {Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes}, series = {Genome Medicine}, volume = {10}, journal = {Genome Medicine}, issn = {1756-994X}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-10538}, pages = {1 -- 12}, year = {2018}, abstract = {Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing method that employs only basic laboratory equipment. We performed a proof-of-concept using mixed mouse hybridoma cells and we also showed that our method can be used for discovery of novel antigen-specific monoclonal antibodies by sequencing human CD19+ B cell IgM and IgG repertoires isolated from peripheral whole blood before and seven days after Td (Tetanus toxoid/Diphtheria toxoid) booster immunization. We anticipate broad applicability of our method for providing insights into adaptive immune responses associated with various diseases, vaccinations, and cancer immunotherapies.}, language = {en} } @article{KagelFrohmeGloekler2019, author = {Kagel, Heike and Frohme, Marcus and Gl{\"o}kler, J{\"o}rn}, title = {Photoacids in biochemical applications}, series = {Journal of Cellular Biotechnology}, volume = {4}, journal = {Journal of Cellular Biotechnology}, number = {1-2}, publisher = {IOS Press}, issn = {2352-3697}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:526-opus4-16123}, pages = {23 -- 30}, year = {2019}, abstract = {BACKGROUND: After excitation with light photoacids can change the pH in a solution by release of a proton. They have been used mostly for excited state proton transfer studies. In this review the general functionality and mechanisms and the subdivision of photoacids is explained. STATE OF THE ART: Different uses of photoacids are described, covering a wide range of various biochemical topics, focusing on biochemical applications. Examples for the introduced subdivisions are covered. CONCLUSIONS AND OUTLOOK: The areas in which photoacids can be employed are diverse. Photoacids have a promising future in biotechnology and biochemistry and should be considered for upcoming applications, especially in non-invasive control of biochemical reactions.}, language = {en} }