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To ensure proper dosage of a drug,analytical quantification of it in biofluid is necessary.Liquid chro-matography mass spectrometry(LC-MS)is the conventional method of choice as it permits accurate identification and quantification.However,it requires expensive instrumentation and is not appropriate for bedside use.Using soluble epoxide hydrolase(sEH)inhibitors(EC5026 and TPPU)as examples,we report development of a nanobody-based enzyme-linked immunosorbent assay(ELISA)for such small molecules and its use to accurately quantify the drug chemicals in human samples.Under optimized conditions,two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL,respectively,and two order of magnitude linear ranges with high precision and accuracy.The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026.In addition,the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency.The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds.These methods could be easily implemented by the bedside,in the field in remote areas or in veterinary practice.This work il-lustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy.At-tributes of nanobody based pharmaceutical assays are discussed.
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Aim To evaluate the potency of anti-D. acutus venom IgY neutralizing the main activities of D. acutus venom.Methods After mixing the different a-mounts of IgY with snake venom and incubating togeth-er,the main activities of snake venom were assayed by biochemical methods.Results The in vitro assays in-dicated that anti-D.acutus venom IgY obviously neu-tralized the activities of PLA2 ,5′-nucleotidase,hyalu-ronidase,metalloprotease and serine proteinase (fi-brinogenase)in D.acutus venom.Mouse experiments showed that the ED50 value of IgY for mouse was 1 131.09 μg.Conclusion Anti-D.acutus venom IgY antibodies have good effects in neutralizing D.acutus venom without the toxicities themselves.
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Objective To study the expression of human papillomavirus ( HPV) and ATM protein in laryngeal squamous cell carcinoma (LSCC), and discuss the correlation among the expression and its prognosis Method The expression of HPV16/18 mRNA was detected by PCR , and the expression of ATM proteins by immunohistochemical method in 63 LSCC specimens and 30 specimens of normal tissue adjacent to cancer. Results The positive expression rates of HPV16 /18 and ATM protein in LSCC group were 39.7% (25 /63) and 41. 3% (26 /63) respectively and those of HPV16 /18 and ATM protein in normal group were 9.5% ( 6/63 ) and 83.3% (25 /30) respectively. There was no correlation between the expression of ATM and HPV16/18 in LSCC. The accumulative 5-year survival rates of HPV16/18 positive group and HPV16/18 negative group in 63 patients were 69.8% and 52.6% respectively ,and there was no significant difference (P> 0.05. The accumulative 5-year survival rates of ATM positive group and ATM negative group in 63 patients were 65.4%and 45.9% respectively and there was no significant difference (P>0.05. Conclusion Both HPV16/18 and ATM are abnormally expressed in LSCC , but there is no correlation between the expression of HPV16/18 and ATM and the expression and its prognosis.
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Objective To investigate effect of the venom peptide liquor on adjunctive arthritis (AA) in mice .Methods The male Kunming mice were randomly divided into 6 groups ,named as the control group ,model group ,positive control group(1 mg/kg dex-amethason) ,wine group(10 mL/kg) ,low dosage of venom peptide liquor group(8 .3 mL/kg) and high dosage of venom peptide liq-uor group(33 .2 mL/kg);the right toes ,ankle diameter and whole body weight were measured at 7-day intervals ;the spleen index , serum level of circulating immune complexes(CIC) were determined after the thirty-fourth day when the mice was put to death .Re-sults The level of CIC in AA mice decreased significantly (P<0 .05) ,the decrease of spleen index and the reduction of toe and an-kle swelling in the low dose group were significant(P<0 .01) .Conclusion The venom peptide liquor exhibited apparent inhibitory effect on AA in mice .
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Purpose To investigate the effect of 5'-nucleotidase from Trimeresurus albolabris venom on platelet aggregations and its mechanism.Methods Piatelet aggregations induced by ADP,AA and PAF,respectively.were measured by turbidimetric method after platelet incubated with 5'-nucleotidase.Results 5'-nucleotidase significantly inhibited platelet aggregations induced by ADP,AA and PAF in a dose-dependent manner.Washed platelet inhibition experiment and the effect of Adenosine,CP/CPK on platelet aggregations showed that platelet aggregations inhibited by 5'-nucleotidase were associated with ADP hydrolysis and adenosine accumulation.The effect of ASA on platelet aggregations showed that the enzyme inhibited platelet aggregations probably by blocking the formation of TXA2.Conclusion 5'-nucleotidase from Trimeresurus albolabris venom inhibited platelet aggregations through ADP hydrolysis and adenosine accumulation,and maybe it is related to blocking the formation of TXA2.