Objective To develop and verify a pre-column derivatization reverse phase-high performance liquid chromatography(RP-HPLC) method for determination of Glycine and Histidine content in recombinant proteins.Methods AccQ Tag-C 18(3.9 mm × 150 mm,4 μm) column was used as chromatographic column,6-aminoquinolyl-N-hydroxysccinimidyl carbamate(AQC) was used as pre-column derivatization reagent,while α-aminobutyric acid as internal standard.AccQTag Eluent A solution,acetonitrile solution and high-purity water were used as mobile phases.The UV detection wavelength was 248 nm,injection volume was 10 μL,flow rate was 1.0 mL/min,and column temperature was 37 ℃.The contents of Glycine and Histidine in samples were determined by the internal standard method,and the specificity,linearity,detection limit,quantitative limit,precision,accuracy and stability of the method were verified.Results The developed method effectively separated Glycine,Histidine and internal standard α-aminobutyric acid with high specificity.The standard curves of Glycine in the range of 2.25～11.25 μg/mL and Histidine in the range of 72.85～364.24 μg/mL showed good linearity,each correlation coefficient(R~2) 0.99.The detection limits were 2.25 μg/mL for Glycine and 18.21 μg/mL for Histidine.The quantitative limits were 4.69 μg/mL for Glycine and 32.86 μg/mL for Histidine.The relative standard deviation(RSD) of 6 replicates with the same concentration of Glycine and Histidine were 4.6% and 5.0%,and the RSD of recovery rate in intermediate precision test was 6.9% and 2.0%,respectively.The content of Glycine was close to the quantitative limit,and the average recoveries of high,medium and low concentrations of samples were within 75.9%～111.7%;The recoveries of Histidine ranged from 88.9% to 97.3%.The RSD of Glycine content and Histidine content was 7.7% and 3.3% respectively at 0,12,18,24,30 and 48 h in the same sample.Conclusion The pre-column derivatization RP-HPLC method has accurate and reliable results with high precision,which might be used for quality control of Glycine and Histidine content in recombinant proteins.

Genetic Engineering ; methods ; trends ; Genetic Therapy ; methods ; trends ; Humans

Anti-Bacterial Agents ; metabolism ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Biological Evolution ; Enterobacteriaceae ; enzymology ; pathogenicity ; Gene Transfer, Horizontal ; Host-Parasite Interactions ; Humans ; beta-Lactam Resistance ; drug effects ; genetics ; beta-Lactamases ; genetics ; metabolism

Genes, Synthetic ; Genetic Engineering ; Genome, Fungal ; Mycoplasma mycoides ; genetics ; Synthetic Biology

Animals ; History, 20th Century ; History, 21st Century ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; history ; virology ; Swine ; virology

Antimalarials ; history ; isolation & purification ; therapeutic use ; Awards and Prizes ; China ; History of Medicine ; History, 20th Century ; Humans ; Malaria ; drug therapy ; Medicine

Colorectal Neoplasms ; diagnosis ; Humans ; Neoplasm Recurrence, Local ; diagnosis ; Neoplastic Stem Cells ; metabolism ; Predictive Value of Tests ; Receptor, EphB2 ; metabolism

This review introduced the application of hippotherapy, as a method of rehabilitation, in the field of children's rehabilitation, the neurological skeletal muscle diseases (stroke, brain traumatic, etc.) and the elderly population. In addition, the neuromuscular activation principle, the core strength principle, and the sensory integration principle were discussed. In order to increase its popularity in the field of rehabilitation, it should combine man-machine interaction with modern physical rehabilitation technology.

BACKGROUND: Brachial plexus root avulsion is the severest type in brachial plexus injuries. It is preganglionic lesion and its prognosis is very bad. At present, the curative effect of traditional neurotization is not satisfactory. Therefore it is necessary to explore the treatment of brachial plexus root avulsion by tissue-engineered neural transplantation. OBJECTIVE: To explore the role and effect of tissue engineering in nerve injury, so as to provide reference for the clinical treatment of brachial plexus root avulsion using tissue engineered neural transplantation. RETRIEVAL STRATEGY: The relevant articles published between January 2001 and October 2007 were retrieved from PubMed and China Journal Full-text Database using the key English words "brachial plexus avulsion" and Chinese words "brachial plexus injury". 410 literatures were retrieved, including 155 English, and 255 Chinese. Focused articles and literatures in the same field published in the near future or on the authoritative journals were included. Repetitive articles and Meta analysis articles were excluded. Finally 34 articles were selected. LITERATURE EVALUATION: Of 34 articles, one was review article, and the others were clinical and experimental studies. DATA SYNTHESIS: Curative effects of traditional nerve transposition and avulsed never reimplantation are limited. Neurotrophic factor, sialidase, chondroitinase ABC, and N-acetyl cysteine are able to protect and promote injured neuron. Bone marrow stem cells and spinal nerve stem cells can differentiate into neuron at lesion site of spinal cord after transplantation. CONCLUSION: Tissue engineered neural transplantation may be a more effective therapy for brachial plexus root avulsion.

Adrenoleukodystrophy ; genetics ; therapy ; Animals ; Color Vision Defects ; genetics ; therapy ; Dependovirus ; Gene Transfer Techniques ; Genetic Therapy ; ethics ; trends ; Hematopoietic Stem Cell Transplantation ; Humans ; Leber Congenital Amaurosis ; genetics ; therapy ; Lentivirus