ABSTRACT
OBJECTIVE To evaluate the bioequivalence and safety of two k inds of Nadroparin calcium injection in healthy Chinese volunteers by subcutaneous injection. METHODS According to the block randomization method ，24 Chinese healthy adult volunteers were included and divided into TR （test preparation-reference preparation ）group and RT （reference preparation-test preparation）group at a ratio of 1∶1. A randomized ，open-labelled，single-dose and two-cycle crossover study was designed ，the fasting subjects of two groups were given test or reference preparation 6 150 AⅩaIU subcutaneously on the first day of each cycle and exchanged in the second cycle ，and the wash-out period was 7 days. The blood samples were collected at different time points before and after administration. The activity of anti-coagulant factor Ⅹa（Anti-Ⅹa）and Anti- Ⅱa in human plasma were determined by chromogenic substrate method ，and the pharmacodynamic parameters were calculated according to the non-atrioventricular model and the bioequivalence was evaluated. The occurrence of adverse events （AEs）was recorded. RESULTS After administration ，the main pharmacodynamic parameters for Anti- Ⅹa activity of test preparation and reference preparation were as follows ：t1/2 were（4.87±1.06） and（4.03±1.00）h，tmax were 4.50（2.00，8.00）and 5.50（2.50，8.00）h，Anti-Ⅹamax were（0.66±0.12）and（0.56±0.11）IU/mL；main pharmacodynamic parameters of Anti- Ⅱa activity of two preparations were as follows ：t1/2 were（3.64±1.60）and（5.74±7.23）h，tmax were 4.00（2.50，8.00）and 4.00（2.00，8.00）h，Anti-Ⅱamax were both （0.10±0.03）IU/mL. The values of 90％confidence interval of geometric mean ratio of Anti- Ⅹamax，AUEC0－t and AUEC 0－∞ were 110.98％-123.50％，112.11％-121.24％and 111.57％-120.00％， respectively. During experiment ，14 subjects reported 19 cases of mild AEs ，among which hematoma ，purpura and maculopapular rash may be related to drugs ；no serious AEs were observed. CONCLUSIONS The domestic Nadroparin calcium injection is bioequivalent to the reference preparation ，and both of them show good safety.
ABSTRACT
Objective To prepare monoclonal antibody (mAb) against recombinant human cardiac troponin I(cTnI).Methods The full-length gene encoding human cardiac troponin I (cTnI) was synthesized chemically and inserted into expression plasmid pBV220 to construct recombinant plasmid p pBV220/cTnI. The recombinant plasmid was transformed into E.coli DH5? which then expressed cTnI. The immunological activity of the expressed cTnI was analyzed by Western blot. Recombinant human cTnI protein was used as antigen to immunize BALB/c mice. Monoclonal anti-bodies against cTnI were prepared by normal hybridoma technology. The relative affinity of mAbs was determined by ELISA. Specificity of mAbs was analyzed by Western blot.Results Human cTnI gene was synthesized and confirmed by DNA sequencing. Positive recombinant clones were identified by restriction enzyme digestion analysis and DNA sequencing. Western blot analysis showed that the cTnI protein could be recognized by an anti- cTnI antibody. Two hybridmas producing antibodies against cTnI were obtained. IgG isotypes of two mAbs were IgG2a and IgG2b. Western blot showed that the antibodies were specific for cTnI. Neutralisation test showed that these mAbs could be evidently neutralized by cTnI.Conclusion The recombinant expression plasmid of cTnI was constructed successfully and expressed in E.coli. The method of EL ISA established to test serum cTnI is to clinically useful. The cTnI mAb which using cTnI as antigen prepared in this paper can be used for cTnI immunoassay in vitro.