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In recent history, alternative approaches to Edman sequencing have been investigated, and to this end, the Association of Biomolecular Resource Facilities (ABRF) Protein Sequencing Research Group (PSRG) initiated studies in 2014 and 2015, looking into bottom-up and top-down N-terminal (Nt) dimethyl derivatization of standard quantities of intact proteins with the aim to determine Nt sequence information. We have expanded this initiative and used low picomole amounts of myoglobin to determine the efficiency of Nt-dimethylation. Application of this approach on protein domains, generated by limited proteolysis of overexpressed proteins, confirms that it is a universal labeling technique and is very sensitive when compared with Edman sequencing. Finally, we compared Edman sequencing and Nt-dimethylation of the same polypeptide fragments; results confirm that there is agreement in the identity of the Nt amino acid sequence between these 2 methods.
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http://dx.doi.org/10.7171/jbt.16-2702-002 | DOI Listing |
Sci Rep
August 2025
Hinge Bio, Inc., Burlingame, CA, 94010, USA.
Recent clinical studies suggest that more potent B cell depleting therapies and targeting more than one B cell antigen may result in improved clinical responses in autoimmune diseases and hematological malignancies. Here we describe an anti-CD19/CD20 bispecific antibody, HB2198, generated using GEM-DIMER™ technology. HB2198 incorporates Fab domains from rituximab and humanized FMC63 (huFMC63) for bivalent binding of both CD19 and CD20 and comprises two enhanced Fc domains to enable powerful effector functions via bivalent binding of Fcγ receptors (FcγR).
View Article and Find Full Text PDFFront Microbiol
June 2025
Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.
Microalgae are promising platforms for recombinant protein production due to their scalability, rapid growth, safety, and sustainability. One strategy to reduce downstream processing costs is to secrete recombinant proteins directly into the culture medium, facilitated by signal peptides (SPs). However, the limited availability of effective SPs has hindered broader applications of this approach in microalgae.
View Article and Find Full Text PDFJ Pharm Biomed Anal
September 2025
School of Life Sciences, Jilin University, Changchun 130012, China; Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun 130012, China. Electronic address:
PEGylation, the covalent attachment of poly(ethylene glycol) (PEG) to biomolecules, is a well-established molecular modification strategy that enhances the pharmacological properties of conjugated proteins by improving stability, reducing immunogenicity, extending half-life, and increasing solubility. Despite its benefits, PEGylated proteins are often heterogeneous mixtures containing positional isoforms that are challenging to eliminate and can significantly impact the biological activity of drugs. This study developed a reversed-phase ultra-performance liquid chromatography (RP-UPLC) method for the separation and quantification of positional isoforms in commercial PEGylated recombinant human growth hormone (PEG-rhGH) at the peptide level.
View Article and Find Full Text PDFBBA Adv
March 2025
Graduate School of NanoBiosciences, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama 236-0027, Japan.
We here report the novel primary structure of a new member in the galectin family, the β-galactoside-binding lectin HOL-30, from the marine sponge , whose full-length sequence was determined thanks to the combination between Edman degradation and transcriptome analysis. The HOL-30 polypeptide is a tandem-repeat dimeric galectin, consisting of 281 amino acids, which includes two carbohydrate recognition domains (CRDs). Unlike most other galectins described in Porifera, HOL-30 did not have a signal peptide sequence for secretion.
View Article and Find Full Text PDFSkelet Muscle
March 2025
Department of Physiology, Nutrition and Biomechanics, The Swedish School of Sport and Health Sciences, Stockholm, Sweden.
Background: Exercising with low muscle glycogen content can improve training adaptation, but the mechanisms underlying the muscular adaptation are still largely unknown. In this study, we measured substrate utilization and cell signaling in different muscle fiber types during exercise and investigated a possible link between these variables.
Methods: Five subjects performed a single leg cycling exercise in the evening (day 1) with the purpose of reducing glycogen stores.