Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective:A camelid natural nanobody library was screened to target S100 calcium-binding protein B (S100B) for obtaining high-affinity and specific nanobodies (Nbs), which provided a molecular basis for the early diagnosis and prognostic treatment of malignant melanoma.Methods:In this study, affinity panning was employed to isolate S100B-specific nanobodies with unique complementarity-determining region 3 (CDR3) sequences from a camelid natural nanobody library. The selected Nbs were expressed in a prokaryotic system and purified via Ni-NTA affinity chromatography. The affinity, specificity, and diagnostic potential of the Nbs were evaluated using enzyme-linked immunosorbent assay (ELISA), western blot, and bio-layer interferometry (BLI).Results:A camelid natural anti-S100B nanobody library with a capacity of 1.91×10 8 CFU was successfully constructed. Affinity panning yielded 30 S100B-specific Nbs, among which Nb107, Nb122, Nb212, and Nb324 with distinct CDR3 sequences were selected for expression. Following Ni-NTA purification, all four anti-S100B Nbs exhibited high purity. Western blot analysis confirmed their ability to recognize recombinant S100B. ELISA and BLI analyses revealed that Nb212 demonstrated high affinity (1.96×10 -11 mol). Additionally, Nb107, Nb122, and Nb212 exhibited broad-spectrum recognition capabilities, binding to various tumor cell lines (Hepa1-6, GL261, 4T1, CT26) as well as murine/human melanoma cells. These Nbs also effectively bound to native murine/human antigens in serum samples from melanoma (A375, B16F10) mouse models. Conclusions:Specific anti-S100B Nbs are successfully screened and expressed, demonstrating not only recognition of native conformational antigens but also broad-spectrum binding and high affinity. These findings highlight their significant potential for developing early diagnostic assays and broad-spectrum targeted vaccines or therapeutics against diverse tumor cells.

Objective:To explore the changing characteristics of self-efficacy, social support, psychological stress in patients with osteoporotic vertebral compression fractures (OVCFs) receiving conservative treatment at different treatment stages and their correlations with the recovery of lower back function.Methods:A total of 116 patients with acute OVCFs who were admitted to the Affiliated Hospital of Medical School, Ningbo University were selected from January 2017 to May 2019. Oswestry Disability Index (ODI) Questionnaire, General Self Efficacy Scale (GSES) , Perceived Social Support Scale (PSSS) and Perceived Stress Scale (PSS) were used to assess the lower back function, self-efficacy, social support and psychological stress of OVCFs patients. Questionnaires were filled out during outpatient reexamination or in-home follow-up, and the evaluation time was at the time of treatment and 1, 3, 6, and 12 months after conservative treatment. Analysis of variance and SNK- q test were used to explore the characteristics of each score over time, and linear regression analysis was used to explore the effects of GSES, PSSS, and PSS on ODI. Results:With the prolongation of treatment time, ODI, GSES, PSSS and PSS scores of OVCFS patients under conservative treatment showed a downward trend, and the scores at 3, 6 and 12 months after treatment were lower than those at 1 month after conservative treatment ( P<0.05) . The scores at 6 and 12 months after conservative treatment were lower than those at 3 months after conservative treatment ( P<0.05) . The scores at 12 months after conservative treatment were lower than that those at 6 months after conservative treatment ( P<0.05) . GSES, PSSS (total score, in-family support, out-of-family support) and PSS were independent influencing factors of ODI at one month after treatment ( P<0.05) . GSES, PSSS (total score, in-family support) and PSS were the influencing factors of ODI at 3 months after treatment ( P<0.05) . Conclusions:The self-efficacy and social support of conservatively treated patients with OVCFs are positively correlated with their lower back function, while psychological stress will reduce lower back function of such patients. It is suggested to add these non-physical factors to the rehabilitation nursing model and take appropriate intervention measures.