Production of Monoclonal Antibodies against Human Trefoil Factor 3 and Development of a Modified-Sandwich ELISA for Detection of Trefoil Factor 3 Homodimer in Saliva

Khummuang, Saichit; Phanphrom, Waraporn; Laopajon, Witida; Kasinrerk, Watchara; Chaiyarit, Ponlatham

doi:10.1186/s12575-017-0064-3

Cited by 5 publications

(3 citation statements)

References 46 publications

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“…Previously, using our generated anti-ζ-globin chain mAbs, an ELISA for detecting ζ-globin chain had been developed [19]. The ELISA in microplate platform, however, has several drawbacks, including time consumption, complicated procedures, the requirement of sophisticated laboratory instrumentation with well-trained technicians and unsuitability for conducting individual tests [13, 14, 16–21]. In this study, we modified our ELISA to an immunostick test platform.…”

Section: Discussionmentioning

confidence: 99%

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

et al. 2019

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Background Couples who carry α-thalassemia-1 deletion are at 25% risk of having a fetus with hemoglobin Bart’s hydrops fetalis. Southeast Asian deletion (−-(SEA)) is the most common type of α-thalassemia 1 among Southeast Asian populations. Thus, identification of the (−-(SEA)) α-thalassemia 1 carrier is necessary for controlling severe α-thalassemia in Southeast Asian countries. Results Using our generated anti ζ-globin chain monoclonal antibodies (mAbs) clones PL2 and PL3, a simple immunostick test for detecting ζ-globin chain presence in whole blood lysates was developed. The procedure of the developed immunostick test was as follows. The immunostick paddles were coated with 50 μg/mL of mAb PL2 as capture mAb, or other control antibodies. The coated immunostick was dipped into cocktail containing tested hemolysate at dilution of 1:500, 0.25 μg/mL biotin-labeled mAb PL3 and horseradish peroxidase-conjugated streptavidin at dilution of 1:1000. The immunostick was then dipped in precipitating substrate and the presence of ζ-globin chain in the tested sample was observed by the naked eye. Upon validation of the developed immunostick test with various types of thalassemia and normal subjects, 100% sensitivity and 82% specificity for detection of the (−-(SEA)) α-thalassemia-1 carriers were achieved. The mAb pre-coated immunostick can be stored at room temperature for at least 20 weeks. Conclusion In this study, a novel simple immunostick test for the screening of (−-(SEA)) α-thalassemia 1 carriers was presented. The developed immunostick test, within a single test, contains both positive and negative internal procedural controls.

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Section: Discussionmentioning

confidence: 99%

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

et al. 2019

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“…For the quantification of salivary TFF3 concentrations, we used our generated monoclonal antibody (mAb) clones 286 and 116 for a modified sandwich enzyme-linked immunosorbent assay (ELISA) technique. 20 To develop the sensitivity, fluorescein isothiocyanate (FITC) and anti-FITC identification methods were used in this sandwich ELISA technique. An antihuman TFF3 mAb clone 286 was used as the plate-coating antibody to capture the peptides in saliva samples.…”

Section: Methodsmentioning

confidence: 99%

Prolonged Suppressive Effects of Periodontitis on Salivary TFF3 Production

Hormdee

Prajaneh

Kampichai³

et al. 2019

Eur J Dent

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Objective As a follow-up to our previous study that demonstrated decreased salivary trefoil factor family 3 (TFF3) peptide levels in chronic periodontitis patients, this current study aimed to observe the effects of nonsurgical periodontal treatment on salivary TFF3 peptides in patients with periodontal diseases. Materials and Methods Eighty-seven volunteers that comprised of 30 individuals with healthy periodontium, 31 with gingivitis, and 26 with chronic periodontitis were considered for the study. Prior to periodontal treatment, a general periodontal examination was performed along with collection of saliva samples from each volunteer. Nonsurgical periodontal treatments were provided to patients with gingivitis and periodontitis. Two weeks post-treatment, saliva samples were recollected, and the periodontal status was re-evaluated. Salivary TFF3 concentrations were measured by enzyme-linked immunosorbent assay. Statistical Analysis Mann–Whitney U test was used when the investigated data were not normally distributed. Chi-squared test was used when dealing with categorical data. Kruskal–Wallis test with post-hoc corrections was used to compare data among the three investigated groups. Two-tailed p < 0.05 was considered as statistically significant. Results Prior to the periodontal treatment, salivary TFF3 concentrations in patients with gingivitis and periodontitis were significantly lower than those with healthy periodontium. Two weeks post-treatment, increased levels of salivary TFF3 were observed in patients with gingivitis, whereas the concentrations decreased in patients with chronic periodontitis. Conclusion This study demonstrated the effects of periodontal disease on the production of salivary TFF3 peptides. Interestingly, nonsurgical periodontal treatment also affected the recovery of salivary TFF3 peptides but varied in their outcomes between gingivitis and periodontitis patients.

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“…About 20 to 80% of human salivary TFF3 exist in a high-molecular-mass form, which represents a TFF3-FCGBP heterodimer (Figure 2A) [17]. The low-molecularmass fractions mainly represent different homodimeric TFF3 forms [17,47]. In the latter, a truncated TFF3 form was also characterized missing the C-terminal phenylalanine residue [17].…”

Section: Potential Role Of Salivary Tff3mentioning

confidence: 99%

Salivary Trefoil Factor Family (TFF) Peptides and Their Roles in Oral and Esophageal Protection: Therapeutic Potential

Hoffmann

2021

IJMS

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Human saliva is a complex body fluid with more than 3000 different identified proteins. Besides rheological and lubricating properties, saliva supports wound healing and acts as an antimicrobial barrier. TFF peptides are secreted from the mucous acini of the major and minor salivary glands and are typical constituents of normal saliva; TFF3 being the predominant peptide compared with TFF1 and TFF2. Only TFF3 is easily detectable by Western blotting. It occurs in two forms, a disulfide-linked homodimer (Mr: 13k) and a high-molecular-mass heterodimer with IgG Fc binding protein (FCGBP). TFF peptides are secretory lectins known for their protective effects in mucous epithelia; the TFF3 dimer probably has wound-healing properties due to its weak motogenic effect. There are multiple indications that FCGBP and TFF3-FCGBP play a key role in the innate immune defense of mucous epithelia. In addition, homodimeric TFF3 interacts in vitro with the salivary agglutinin DMBT1gp340. Here, the protective roles of TFF peptides, FCGBP, and DMBT1gp340 in saliva are discussed. TFF peptides are also used to reduce radiotherapy- or chemotherapy-induced oral mucositis. Thus, TFF peptides, FCGBP, and DMBT1gp340 are promising candidates for better formulations of artificial saliva, particularly improving wound healing and antimicrobial effects even in the esophagus.

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Production of Monoclonal Antibodies against Human Trefoil Factor 3 and Development of a Modified-Sandwich ELISA for Detection of Trefoil Factor 3 Homodimer in Saliva

Cited by 5 publications

References 46 publications

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Immunostick Test for Detecting ζ-Globin Chains and Screening of the Southeast Asian α-Thalassemia 1 Deletion

Prolonged Suppressive Effects of Periodontitis on Salivary TFF3 Production

Salivary Trefoil Factor Family (TFF) Peptides and Their Roles in Oral and Esophageal Protection: Therapeutic Potential

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