Solid-Phase Enzyme Immunoassay for the Detection of HMG Nonhistone Proteins in Their Native Structure

Lepp, Wolfgang A.; Martı́nez, Paz

doi:10.1080/01971528908053252

Cited by 7 publications

(2 citation statements)

References 16 publications

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“…However, currently, the only commercially available HMGB1 ELISA kit is from Shino-Test Corporation in Japan, first reported in 2006 (Yamada et al, 2006). In addition to ELISA (Lepp and Martinez, 1989; Urbonaviciute et al, 2007; Wahamaa et al, 2007; Yamada et al, 2003), several new techniques (e.g., DNA nanostructure-based assay) have been proposed to test HMGB1 concentration in serum or supernatants (Gaillard et al, 2008). Immunohistochemical staining is widely used in the assessment of HMGB1 expression and localization in tissues.…”

Section: Hmgb1 Measurements and Significance In Clinical Practicementioning

confidence: 99%

HMGB1 in health and disease

Kang

Chen

Zhang

et al. 2014

Molecular Aspects of Medicine

823

828

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Complex genetic and physiological variations as well as environmental factors that drive emergence of chromosomal instability, development of unscheduled cell death, skewed differentiation, and altered metabolism are central to the pathogenesis of human diseases and disorders. Understanding the molecular bases for these processes is important for the development of new diagnostic biomarkers, and for identifying new therapeutic targets. In 1973, a group of non-histone nuclear proteins with high electrophoretic mobility was discovered and termed High-Mobility Group (HMG) proteins. The HMG proteins include three superfamilies termed HMGB, HMGN, and HMGA. High-mobility group box 1 (HMGB1), the most abundant and well-studied HMG protein, senses and coordinates the cellular stress response and plays a critical role not only inside of the cell as a DNA chaperone, chromosome guardian, autophagy sustainer, and protector from apoptotic cell death, but also outside the cell as the prototypic damage associated molecular pattern molecule (DAMP). This DAMP, in conjunction with other factors, thus has cytokine, chemokine, and growth factor activity, orchestrating the inflammatory and immune response. All of these characteristics make HMGB1 a critical molecular target in multiple human diseases including infectious diseases, ischemia, immune disorders, neurodegenerative diseases, metabolic disorders, and cancer. Indeed, a number of emergent strategies have been used to inhibit HMGB1 expression, release, and activity in vitro and in vivo. These include antibodies, peptide inhbitiors, RNAi, anti-coagulants, endogenous hormones, various chemical compounds, HMGB1-receptor and signaling pathway inhibition, artificial DNAs, physical strategies including vagus nerve stimulation and other surgical approaches. Future work further investigating the details of HMGB1 localizationtion, structure, post-translational modification, and identifccation of additional partners will undoubtedly uncover additional secrets regarding HMGB1’s multiple functions.

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Section: Hmgb1 Measurements and Significance In Clinical Practicementioning

confidence: 99%

HMGB1 in health and disease

Kang

Chen

Zhang

et al. 2014

Molecular Aspects of Medicine

823

828

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“…The ELISA procedure employed was essentially as described previously [26]. Briefly, 10 jIg of purified AWN-1 spermadhesin in 100 pIl of 50 mM Na 2 CO 3 /NaHCO 3 , pH 9.6, was coated to flat-bottom polystyrene plates overnight at 4°C.…”

Section: Elisa For the Selection Of Monoclonal Antibodiesmentioning

confidence: 99%

Monoclonal Antibodies against Boar Sperm Zona Pellucida-Binding Protein AWN-1. Characterization of a Continuous Antigenic Determinant and Immunolocalization Of AWN Epitopes in Inseminated Sows1

Calvete

Ensslin

Mburu

et al. 1997

Biology of Reproduction

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Boar spermadhesin AWN-1 is a sperm surface-associated 14.7-kDa lectin and a major protein of porcine seminal plasma. AWN-1 binds to beta-galactosides and to porcine zona pellucida glycoproteins, suggesting that this protein might play a role in the primary binding of spermatozoa to the egg's external glycoprotein matrix. We have produced a collection of murine monoclonal antibodies against purified AWN-1. Five monoclonal antibodies recognized sequential antigenic determinants. All these epitopes were located at the C-terminal region of AWN-1 (residues 109-123) by competitive ELISA using overlapping synthetic peptides that cover the complete 133 amino acid sequence of the lectin. In a structural model of spermadhesin AWN-1, the polypeptide stretch 109-123 is fully solvent-exposed, providing a reasonable explanation for its high immunogenicity. In addition to epitope mapping, we have employed anti-AWN monoclonal antibodies for immunolocalization of the protein in the genital tract of inseminated sows. Clusters of AWN epitopes were occasionally found attached to the epithelium of the uterotubal junction and the adjacent lower isthmus. However, neither AWN-1 nor other seminal plasma proteins were found in the isthmic fluid collected 10-26 h after insemination. These results suggest that the whole amount of seminal plasma proteins are absorbed by the epithelium of the female genital tract, supporting the claim that removal of seminal plasma components from spermatozoa might be a major event in both in vitro and in vivo sperm capacitation.

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