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Quenchbody (Q-body) is a unique, reagentless, fluorescent antibody whose fluorescent intensity increases in an antigen-concentration-dependent manner. Q-body-based homogeneous immunoassay is superior to conventional immunoassays as it does not require multiple immobilization, reaction, and washing steps. In fact, simply mixing the Q-body and the sample containing the antigen enables the detection of the target antigen. To date, various Q-bodies have been developed to detect biomarkers of interest, including haptens, peptides, proteins, and cells. This review sought to describe the principle of Q-body-based immunoassay and the use of Q-body for various immunoassays. In particular, the Q-bodies were classified from a structural perspective to provide useful information for designing Q-bodies with an appropriate objective. [ABSTRACT FROM AUTHOR]

Factor VIII (F8) is a blood coagulation protein prearranged in six domains, and its deficiency causes hemophilia A. To fashion functional F8 therapeutics, development of a recombinant F8 (rF8) domain is essential not only for F8 substitution, but also to decipher the F8-related mechanisms. In this study, we generated Glutathione S-transferase (GST)-conjugated recombinant A2 and A3 domains of F8 using Escherichia coli. The high growth rate and economically advantageous protein production system in terms of inexpensive reagents and materials in E. coli cells facilitated the completion of entire process from protein expression to purification in 3–4 days with low production cost. Subsequent assessment of these purified proteins using enzyme-linked immunosorbent assay (ELISA) and antibodies against F8 revealed enhanced detection of rF8-A2 or rF8-A3 in a concentration dependent manner, indicating the presence of the antibody-binding epitopes in these proteins. Furthermore, these proteins are suitable for generating novel antibodies against the F8 domain and F8 domain-capturing affinity columns by enabling their conjugation to GST-capturing beads. Additionally, the recombinant F8 domains produced herein can be used for various studies, which include investigating the explicit roles of the F8 domain in the coagulation process, with domain-specific binding partners, and antibodies. [ABSTRACT FROM AUTHOR]

Introduction The lack of genetic diversity in modern crops is a serious problem owing to the concurrent vulnerabilities of crop plants to biotic and abiotic stresses (Hammer et al. 2003). [...], Single nucleotide polymorphisms (SNPs) derived from both nuclear and cytoplasmic DNA sequences were developed to identify distinct species of Capsicum. Species identification was achieved by detecting allelic variations of these type of markers via high resolution melting analysis (HRM). We used the HRM polymorphisms of COSII markers and the Waxy gene from the nuclear sequence, in addition to the intergenic spacer between trnL and trnF from cytoplasmic DNA as our SNP markers. A total of 31 accessions of Capsicum, representing six species, were analyzed using this method. As single markers were insufficient for identifying Capsicum species, combinations of all markers unambiguously identified all six. A phylogeny based on the SNP markers was consistent with the current taxonomy of Capsicum species. These observations demonstrate that the markers developed in this study are useful for rapid identification of new germplasm for management of Capsicum species. Key words: germplasm, Capsicum, DNA barcoding, single nucleotide polymorphism (SNP), high resolution melting (HRM). Des polymorphismes mononucleotidiques (SNP) deriveis tant de sequences nucleaires que cytoplasmiques ont ete developpes afin d'identifier diverses especes du genre Capsicum. L'identification des especes a ete realisee en detectant des variations alleliques chez ces marqueurs par analyse de fusion a haute resolution (HRM pour << high resolution melting >>). Les auteurs ont etudie le polymorphisme HRM chez les marqueurs COSII et le gene Waxy pour le genome nucleaire en plus des espaceurs intergeiniques entre trnL et trnF au sein de l'ADN cytoplasmique. Au total, 31 accessions du genre Capsicum, representant six especes, ont ete analysees a l'aide de cette methode. Bien que les marqueurs individuels aient ete; insuffisants pour identifier les especes, des combinaisons de ces marqueurs ont permis d'identifier sans ambiguite les six especes. Une phylogenie basee sur les marqueurs SNP etait conforme a la taxonomie actuelle pour les especes du genre Capsicum. Ces observations montrent que les marqueurs developpeis au cours de ce travail sont utiles pour l'identification rapide et la gestion des nouvelles ressources genetiques chez le genre Capsicum. Mots-cles: germoplasme, Capsicum, code barre genetique, polymorphisme mononucleotidique (SNP), fusion a haute resolution (HRM). [Traduit par la Redaction]