المؤلفون: Trina Mouchahoir, John E. Schiel, Rich Rogers, Alan Heckert, Benjamin J. Place, Aaron Ammerman, Xiaoxiao Li, Tom Robinson, Brian Schmidt, Chris M. Chumsae, Xinbi Li, Anton V. Manuilov, Bo Yan, Gregory O. Staples, Da Ren, Alexander J. Veach, Dongdong Wang, Wael Yared, Zoran Sosic, Yan Wang, Li Zang, Anthony M. Leone, Peiran Liu, Richard Ludwig, Li Tao, Wei Wu, Ahmet Cansizoglu, Andrew Hanneman, Greg W. Adams, Irina Perdivara, Hunter Walker, Margo Wilson, Arnd Brandenburg, Nick DeGraan-Weber, Stefano Gotta, Joe Shambaugh, Melissa Alvarez, X. Christopher Yu, Li Cao, Chun Shao, Andrew Mahan, Hirsh Nanda, Kristen Nields, Nancy Nightlinger, Ben Niu, Jihong Wang, Wei Xu, Gabriella Leo, Nunzio Sepe, Yan-Hui Liu, Bhumit A. Patel, Douglas Richardson, Yi Wang, Daniela Tizabi, Oleg V. Borisov, Yali Lu, Ernest L. Maynard, Albrecht Gruhler, Kim F. Haselmann, Thomas N. Krogh, Carsten P. Sönksen, Simon Letarte, Sean Shen, Kristin Boggio, Keith Johnson, Wenqin Ni, Himakshi Patel, David Ripley, Jason C. Rouse, Ying Zhang, Carly Daniels, Andrew Dawdy, Olga Friese, Thomas W. Powers, Justin B. Sperry, Josh Woods, Eric Carlson, K. Ilker Sen, St John Skilton, Michelle Busch, Anders Lund, Martha Stapels, Xu Guo, Sibylle Heidelberger, Harini Kaluarachchi, Sean McCarthy, John Kim, Jing Zhen, Ying Zhou, Sarah Rogstad, Xiaoshi Wang, Jing Fang, Weibin Chen, Ying Qing Yu, John G. Hoogerheide, Rebecca Scott, Hua Yuan

المصدر: Journal of the American Society for Mass Spectrometry. 33:1659-1677

مصطلحات موضوعية: Quality Control, Benchmarking, Structural Biology, Proteins, Peptide Mapping, Mass Spectrometry, Spectroscopy

الوصف: The multi-attribute method (MAM) was conceived as a single assay to potentially replace multiple single-attribute assays that have long been used in process development and quality control (QC) for protein therapeutics. MAM is rooted in traditional peptide mapping methods; it leverages mass spectrometry (MS) detection for confident identification and quantitation of many types of protein attributes that may be targeted for monitoring. While MAM has been widely explored across the industry, it has yet to gain a strong foothold within QC laboratories as a replacement method for established orthogonal platforms. Members of the MAM consortium recently undertook an interlaboratory study to evaluate the industry-wide status of MAM. Here we present the results of this study as they pertain to the targeted attribute analytics component of MAM, including investigation into the sources of variability between laboratories and comparison of MAM data to orthogonal methods. These results are made available with an eye toward aiding the community in further optimizing the method to enable its more frequent use in the QC environment.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8fbb68de4c53ee4678b9e74066d51588Test
https://doi.org/10.1021/jasms.2c00129Test

المؤلفون: Martin Palmer, Peter Kresten Nielsen, Keith Richardson, Jakub Ujma, Kevin Giles, Nick Tomczyk, Kim F. Haselmann

المصدر: Analytical Chemistry. 93:16379-16384

مصطلحات موضوعية: chemistry.chemical_classification, Biological Products, Chromatography, Chemistry, Ion-mobility spectrometry, High resolution, Peptide, Mass spectrometry, Mass Spectrometry, Analytical Chemistry, Biopharmaceutical industry, Ion Mobility Spectrometry, Posttranslational modification, Isobaric process, Peptides, Chromatography, Liquid

الوصف: The identification and localization of isomeric peptide modifications is a critical requirement of the biopharmaceutical industry. Despite the ability of liquid chromatography-mass spectrometry to identify many of the common post translational modifications, the identification of isobaric or racemized peptides is confounded by modern mass spectrometry-based techniques. Here, we present a novel approach combining liquid chromatography with a high-resolution ion mobility mass spectrometry system to differentiate peptide and peptide fragments based upon their mobility and mass.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b0725117725e9a72c37b09fc81021231Test
https://doi.org/10.1021/acs.analchem.1c02834Test

المؤلفون: Kim F. Haselmann, Simon K Gammelgaard, Steffen B. Petersen, Peter Kresten Nielsen

المصدر: ACS Omega, Vol 5, Iss 14, Pp 7962-7968 (2020)
Gammelgaard, S K, Petersen, S B, Haselmann, K F & Nielsen, P K 2020, ' Direct Ultraviolet Laser-Induced Reduction of Disulfide Bonds in Insulin and Vasopressin ', ACS Omega, vol. 5, no. 14, pp. 7962-7968 . https://doi.org/10.1021/acsomega.9b04375Test
ACS Omega

مصطلحات موضوعية: Vasopressin, Arginine, Chemistry, General Chemical Engineering, General Chemistry, Mass spectrometry, Photochemistry, medicine.disease_cause, Article, Electron transfer, Excited state, medicine, Irradiation, Spectroscopy, QD1-999, Ultraviolet

الوصف: Ultraviolet (UV) light has been shown to induce reduction of disulfide bonds in proteins in solution. The photoreduction is proposed to be a result of electron donation from excited Tyr or Trp residues. In this work, a powerful UV femtosecond laser was used to generate photoreduced products, while the hypothesis of Tyr/Trp mediation was studied with spectroscopy and mass spectrometry. With limited irradiation times of 3 min or less at 280 nm, the laser-induced reduction in arginine vasopressin and human insulin led to significant yields of ∼3% stable reduced product. The photogenerated thiols required acidic pH for stabilization, while neutral pH primarily caused scrambling and trisulfide formation. Interestingly, there was no direct evidence that Tyr/Trp mediation was a required criterion for the photoreduction of disulfide bonds. Intermolecular electron transfer remained a possibility for insulin but was ruled out for vasopressin. We propose that an additional mechanism should be increasingly considered in UV light-induced reduction of disulfide bonds in solution, in which a single UV photon is directly absorbed by the disulfide bond.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::39366b62321b6403142eb1508c953529Test
https://doaj.org/article/02aab33642364ceaaf6a8e59a6d6b489Test

المؤلفون: Peter Kresten Nielsen, Simon K Gammelgaard, Steffen B. Petersen, Kim F. Haselmann

المصدر: Gammelgaard, S K, Petersen, S B, Haselmann, K F & Nielsen, P K 2021, ' Characterization of Insulin Dimers by Top-Down Mass Spectrometry ', Journal of The American Society for Mass Spectrometry, vol. 32, no. 8, pp. 1910–1918 . https://doi.org/10.1021/jasms.0c00257Test

مصطلحات موضوعية: Stereochemistry, Ultraviolet Rays, Dimer, Iron, Phenylalanine, Peptides and proteins, Mass spectrometry, medicine.disease_cause, Dissociation (chemistry), Mass Spectrometry, Workflow, chemistry.chemical_compound, Fragmentation, Structural Biology, medicine, Side chain, Humans, Insulin, Cysteine, Nucleic acid structure, Spectroscopy, Monomers, Photodissociation, Monomer, chemistry, Covalent bond, Oligomers, Tyrosine, Protein Multimerization, Ultraviolet

الوصف: High-molecular weight products (HMWP) are an important critical quality attribute in research and development of insulin biopharmaceuticals. We here demonstrate on two case studies of covalent insulin dimers, induced by Fe2+ incubation or ultraviolet (UV) light stress, that de novo characterization in top-down mass spectrometry (MS) workflows can identify cross-link types and sites. On the MS2 level, electron-transfer/higher-energy collision dissociation (EThcD) efficiently cleaved the interchain disulfide bonds in the dimers to reveal cross-link connectivities between chains. The combined utilization of EThcD and 213 nm ultraviolet photodissociation (UVPD) facilitated identification of the chemical composition of the cross-links. Identification of cross-link sites between chains at residue level was achievable for both dimers with MS3 analysis of MS2 fragments cleaved at the cross-link or additionally the interchain disulfide bonds. UVPD provided identification of cross-link sites in the Fe2+-induced dimer without MS3, while cross-link site identification with MS2 was not possible for the UV light-induced dimer. Thus, using varied multistage approaches, it was discovered that in the UV light-induced dimer, Tyr14 of the A-chain participated in an -O-S- cross-link in which the sulfur was derived either from Cys7 or Cys19 of the B-chain. In the Fe2+-induced dimer, Phe1 from both B-chains were cross-linked through a -CH2-. The UV chromophoric side chain of Phe1 was indicated in the cross-link, explaining why UVPD-MS2 was effective in fragmenting the cross-link and nearby backbone bonds. Our results demonstrated that higher-energy collisional dissociation (HCD), EThcD, and UVPD combined with MS3 were powerful tools for direct de novo characterization of cross-linked insulin dimers.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ceaed5061186ce873ad5babb41b04f58Test
https://pubmed.ncbi.nlm.nih.gov/33084334Test

المؤلفون: Sam Hughes, Nicolas L. Young, David Clarke, Frank Sobott, Anja Resemann, Kristina Srzentić, Kim F. Haselmann, Detlev Suckau, Dina L. Bai, Young Ah Goo, Christian Malosse, Mathieu Dupré, Sylvester M. Greer, Neil R. Quebbemann, Ziqing Lin, Jared B. Shaw, Alain Beck, Yury O. Tsybin, Stuart Pengelley, Timothy K. Toby, Lidong He, Paul O. Danis, Henrique S. Seckler, Robert Anthony D’Ippolito, Natalia Gasilova, Jeffrey N. Agar, Donald F. Hunt, Jennifer S. Brodbelt, Matthew V. Holt, Simone Nicolardi, Joseph A. Loo, Yuri E. M. van der Burgt, Nicholas D. Schmitt, Laure Menin, Robert J. Millikin, Ljiljana Paša-Tolić, David R. Goodlett, Mowei Zhou, Joshua D. Hinkle, Michael R. Shortreed, Christopher L. Hendrickson, Wendy Sandoval, Richa Sarin, Jeffrey Shabanowitz, Alan G. Marshall, Anton N. Kozhinov, Neil L. Kelleher, Chad R. Weisbrod, Lissa C. Anderson, Julia Chamot-Rooke, Yunqiu Chen, John R. Yates, Luca Fornelli, Konstantin O. Nagornov, Lloyd M. Smith, Sneha Chatterjee, Ying Ge, Sung Hwan Yoon, Jolene K. Diedrich, Norelle C. Wildburger

المساهمون: Centre de Recherche Pierre Fabre (Centre de R&D Pierre Fabre), PIERRE FABRE, Institut Pasteur [Paris], Institut Pasteur [Paris] (IP)

المصدر: Journal of The American Society for Mass Spectrometry
Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), 2020, 31 (9), pp.1783-1802. ⟨10.1021/jasms.0c00036⟩
Journal of the American Society for Mass Spectrometry
Journal of the American Society for Mass Spectrometry, vol 31, iss 9
Journal of The American Society for Mass Spectrometry, 31(9), 1783-1802. AMER CHEMICAL SOC
J Am Soc Mass Spectrom
Journal of The American Society for Mass Spectrometry, 2020, 31 (9), pp.1783-1802. ⟨10.1021/jasms.0c00036⟩

مصطلحات موضوعية: Proteomics, medicine.drug_class, [SDV]Life Sciences [q-bio], Computational biology, 010402 general chemistry, Tandem mass spectrometry, Monoclonal antibody, Mass spectrometry, 01 natural sciences, Article, Antibodies, Mass Spectrometry, FTMS, Analytical Chemistry, Mice, Medicinal and Biomolecular Chemistry, Structural Biology, Monoclonal, intact mass measurement, tandem mass spectrometry, medicine, MS/MS, Animals, Humans, Biology, Spectroscopy, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Chemistry, 010401 analytical chemistry, Therapeutic protein, Antibodies, Monoclonal, Complementarity Determining Regions, 0104 chemical sciences, 3. Good health, Fourier transform mass spectrometry, Ion dissociation, glycoform, Therapeutic antibody, Biotechnology, Physical Chemistry (incl. Structural)

الوصف: International audience; The Consortium for Top-Down Proteomics (www.topdownproteomics.org) launched the present study to assess the current state of top-down mass spectrometry (TD MS) and middle-down mass spectrometry (MD MS) for characterizing monoclonal antibody (mAb) primary structures, including their modifications. To meet the needs of the rapidly growing therapeutic antibody market, it is important to develop analytical strategies to characterize the heterogeneity of a therapeutic product's primary structure accurately and reproducibly. The major objective of the present study is to determine whether current TD/MD MS technologies and protocols can add value to the more commonly employed bottom-up (BU) approaches with regard to confirming protein integrity, sequencing variable domains, avoiding artifacts, and revealing modifications and their locations. We also aim to gather information on the common TD/MD MS methods and practices in the field. A panel of three mAbs was selected and centrally provided to 20 laboratories worldwide for the analysis: Sigma mAb standard (SiLuLite), NIST mAb standard, and the therapeutic mAb Herceptin (trastuzumab). Various MS instrument platforms and ion dissociation techniques were employed. The present study confirms that TD/MD MS tools are available in laboratories worldwide and provide complementary information to the BU approach that can be crucial for comprehensive mAb characterization. The current limitations, as well as possible solutions to overcome them, are also outlined. A primary limitation revealed by the results of the present study is that the expert knowledge in both experiment and data analysis is indispensable to practice TD/MD MS.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ce93ac618ef03bef2381fe00d09bf9cfTest
https://hdl.handle.net/1887/3182316Test

المؤلفون: Christian N. Cramer, Kim F. Haselmann, Jesper V. Olsen, Peter Kresten Nielsen, Christian D. Kelstrup

المصدر: Analytical Chemistry. 90:8202-8210

مصطلحات موضوعية: Models, Molecular, 0301 basic medicine, Mass spectrometry, 01 natural sciences, Protein Structure, Secondary, Workflow, Analytical Chemistry, Ion, 03 medical and health sciences, Protein structure, Tandem Mass Spectrometry, Transforming Growth Factor beta, Liquid chromatography–mass spectrometry, Amino Acid Sequence, Disulfides, Peptide sequence, Chromatography, Chemistry, 010401 analytical chemistry, Proteolytic enzymes, Disulfide bond, 0104 chemical sciences, 030104 developmental biology, Chromatography, Liquid, Peptide Hydrolases, Cysteine

الوصف: Disulfide bond mapping is a critical task in protein characterization as protein stability, structure, and function is dependent on correct cysteine connectivities. Mass spectrometry (MS) is the method of choice for this, providing fast and accurate characterization of simple disulfide bonds. Disulfide mapping by liquid chromatography tandem mass spectrometry (LC-MS/MS) is performed by identifying disulfide-bonded partner peptides following proteolytic digestion. With the recently introduced ability to assign complex disulfide patterns by online postcolumn partial disulfide reduction by in-source reduction (ISR) in a LC-ISR-MS/MS methodology, the main challenge is data analysis to ensure detection of both expected and unexpected disulfide species. In this study, we introduced a workflow for confident and unbiased mapping of complex disulfide bonds using the powerful combination of extracted ion chromatograms (XICs) of LC-ISR-MS/MS data. With postcolumn partial reduction, identical LC retention times of intact disulfide-bonded species, their constituting free peptides, and partially reduced variants were observed. Subsequent selective MS/MS fragmentation of all reduction products allowed confident identification of free cysteine-containing peptides using a classical shotgun proteomics database search. Matching XICs of the identified cysteine-containing peptides allowed identification of both predicted and unpredicted disulfide species, including unforeseen proteolytic specificities, missed cleavage sites, scrambled disulfide variants, and the presence of disulfide-entangled complexes. Applying this workflow, we successfully mapped the complex disulfide bonds of tertiapin and the epidermal growth factor (EGF) family members transforming growth factor α (TGFα) and EGF. In addition, we were able to characterize the disulfide patterns of the special disulfide fold of the TGFβ superfamily in an all-online methodology.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bf57fc88dbb50b5f3413cd783f0ed748Test
https://doi.org/10.1021/acs.analchem.8b01603Test

المؤلفون: Jesper V. Olsen, Christian N. Cramer, Peter Kresten Nielsen, Kim F. Haselmann, Christian D. Kelstrup

المصدر: Analytical Chemistry. 89:5949-5957

مصطلحات موضوعية: 0301 basic medicine, chemistry.chemical_classification, Stereochemistry, Electrospray ionization, 010401 analytical chemistry, Disulfide bond, Peptide, Mass spectrometry, 01 natural sciences, 0104 chemical sciences, Analytical Chemistry, 03 medical and health sciences, 030104 developmental biology, Reduction procedure, chemistry, Disulfide bonding, Organic chemistry, Data dependent, Cysteine

الوصف: Mapping of disulfide bonds is an essential part of protein characterization to ensure correct cysteine pairings. For this, mass spectrometry (MS) is the most widely used technique due to fast and accurate characterization. However, MS-based disulfide mapping is challenged when multiple disulfide bonds are present in complicated patterns. This includes the presence of disulfide bonds in nested patterns and closely spaced cysteines. Unambiguous mapping of such disulfide bonds typically requires advanced MS approaches. In this study, we exploited in-source reduction (ISR) of disulfide bonds during the electrospray ionization process to facilitate disulfide bond assignments. We successfully developed a LC-ISR-MS/MS methodology to use as an online and fully automated partial reduction procedure. Postcolumn partial reduction by ISR provided fast and easy identification of peptides involved in disulfide bonding from nonreduced proteolytic digests, due to the concurrent detection of disulfide-containing peptide species and their composing free peptides. Most importantly, intermediate partially reduced species containing only a single disulfide bond were also generated, from which unambiguous assignment of individual disulfide bonds could be done in species containing closely spaced disulfide bonds. The strength of this methodology was demonstrated by complete mapping of all four disulfide bonds in lysozyme and all 17 disulfide bonds in human serum albumin, including nested disulfide bonds and motifs of adjacent cysteine residues.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::35ddc8881b12f1be2da65bcc09885f86Test
https://doi.org/10.1021/acs.analchem.7b00424Test

المؤلفون: Nick Tomczyk, Kim F. Haselmann, Jeffery Mark Brown, Christian N. Cramer, Peter Kresten Nielsen

المصدر: Journal of the American Society for Mass Spectrometry. 28:384-388

مصطلحات موضوعية: 0301 basic medicine, Collision-induced dissociation, Chemistry, 010401 analytical chemistry, Analytical chemistry, Tandem mass spectrometry, Mass spectrometry, 01 natural sciences, Spectral line, 0104 chemical sciences, Ion, Electron-transfer dissociation, 03 medical and health sciences, 030104 developmental biology, Fragmentation (mass spectrometry), Structural Biology, Quadrupole time of flight, Spectroscopy

الوصف: Data-independent mass spectral acquisition is particularly powerful when combined with ultra-performance liquid chromatography (LC) that provides excellent separation of most components present in a given sample. Data-independent analysis (DIA) consists of alternating full MS scans and scans with fragmentation of all ions within a selected m/z range, providing precursor masses and structure information, respectively. Fragmentation spectra are acquired either by sequential isolation and fragmentation of sliding m/z ranges or fragmenting all ions entering the MS instrument with no ion isolation, termed broadband DIA. Previously, broadband DIA has only been possible using collision induced dissociation (CID). Here, we report the use of electron transfer dissociation (ETD) as the fragmentation technique in broadband DIA instead of traditional collision induced dissociation (CID) during MSE. In this approach, which we refer to as MSETD, we implement the inherent benefits provided by ETD, such as discrimination of leucine and isoleucine, in a DIA setup. The combination of DIA analysis and ETD fragmentation with supplemental CID energy provides a powerful platform to obtain information on all precursors and their sequence from a single experiment. Graphical Abstract ᅟ.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::c71610165647e1a190d1aa826c1f8e2cTest
https://doi.org/10.1007/s13361-016-1538-2Test

المؤلفون: Kasper D. Rand, Jeffery Mark Brown, Ulrik H. Mistarz, Kim F. Haselmann

المصدر: Structure. 24:310-318

مصطلحات موضوعية: Models, Molecular, 0301 basic medicine, Spectrometry, Mass, Electrospray Ionization, Protein Conformation, Electrospray ionization, Analytical chemistry, Mass spectrometry, 01 natural sciences, 03 medical and health sciences, Protein structure, Structural Biology, Binding site, Molecular Biology, Conformational isomerism, Binding Sites, Chemistry, 010401 analytical chemistry, Deuterium Exchange Measurement, 0104 chemical sciences, Crystallography, 030104 developmental biology, Deuterium, Multiprotein Complexes, Hydrogen–deuterium exchange, Protein Binding

الوصف: SummaryFast gas-phase hydrogen/deuterium exchange mediated by ND3 gas and measured by mass spectrometry (gas-phase HDX-MS) is a largely unharnessed, fast, and sensitive method for probing primary- and higher-order polypeptide structure. Labeling of heteroatom-bound non-amide hydrogens in a sub-millisecond time span after electrospray ionization by ND3 gas can provide structural insights into protein conformers present in solution. Here, we have explored the use of gas-phase HDX-MS for probing the higher-order structure and binding interfaces of protein complexes originating from native solution conditions. Lysozyme ions bound by an oligosaccharide incorporated less deuterium than the unbound ion. Similarly, trypsin ions showed reduced deuterium uptake when bound by the peptide ligand vasopressin. Our results are in good agreement with crystal structures of the native protein complexes, and illustrate that gas-phase HDX-MS can provide a sensitive and simple approach to measure the number of heteroatom-bound non-amide side-chain hydrogens involved in the binding interface of biologically relevant protein complexes.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3876dcc1aee60af009b295be1d4c9e02Test
https://doi.org/10.1016/j.str.2015.11.013Test

المؤلفون: Peter Kresten Nielsen, Jesper V. Olsen, Kim F. Haselmann, Christian N. Cramer

المصدر: Analytical Chemistry. 88:1585-1592

مصطلحات موضوعية: Disulfide Linkage, Chemistry, 010401 analytical chemistry, Electrochemical Techniques, 010402 general chemistry, Mass spectrometry, Human serum albumin, 01 natural sciences, Combinatorial chemistry, Mass Spectrometry, 0104 chemical sciences, Analytical Chemistry, Protein sequencing, Protein structure, Covalent bond, medicine, Humans, Insulin, Muramidase, Disulfides, Oxidation-Reduction, Serum Albumin, Cysteine, medicine.drug

الوصف: Unravelling of disulfide linkage patterns is a crucial part of protein characterization, whether it is for a previously uncharacterized protein in basic research or a recombinant pharmaceutical protein. In the biopharmaceutical industry, elucidation of the cysteine connectivities is a necessity to avoid disulfide scrambled and incorrectly folded forms in the final product. Mass spectrometry (MS) is a highly utilized analytical tool for this due to fast and accurate characterization. However, disulfide bonds being an additional covalent bond in the protein structure represent a challenge in protein sequencing by tandem MS (MS/MS). Electrochemical (EC) reduction of disulfide bonds has recently been demonstrated to provide efficient reduction efficiencies, significantly enhancing sequence coverages in online coupling with MS characterization. In this study, the potential use of EC disulfide reduction in combination with MS characterization for disulfide mapping was assessed. We employed two approaches based on (1) the high flexibility and instant information about the degree of reduction in infusion EC-MS to generate partially reduced species on the intact protein level and (2) the preserved link between parent disulfide-linked fragments and free reduced peptides in an LC-EC-MS platform of nonreduced proteolytic protein digestions. Here we report the successful use of EC as a partial reduction approach in mapping of disulfide bonds of intact human insulin (HI) and lysozyme. In addition, we established a LC-EC-MS platform advantageous in disulfide characterization of complex and highly disulfide-bonded proteins such as human serum albumin (HSA) by online EC reduction of nonreduced proteolytic digestions.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7f2803a96e8f184e8867b61594da3d94Test
https://doi.org/10.1021/acs.analchem.5b03148Test