Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was se...Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin （LCA）-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay （RIA）. Results： Circulating AFP was separated into three peaks （AFP-1, AFP-2, and AFP-3） by LCA-affinity chromatography. Dunng the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HSAFP （AFP-3） from sera of HCC patients was eluted clearly on the LCA-sepharose gel mini-column with a solution containing α-methyI-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity （92.7%） and specificity （88.2%）. Conclusion： The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.展开更多
A new kind of electrophoretic affinity chromatography (EAC) for bioseparation was proposed. Separation by EAC was conducted in a multicompartment electrolyzer in which the affinity gel media were packed in one of the ...A new kind of electrophoretic affinity chromatography (EAC) for bioseparation was proposed. Separation by EAC was conducted in a multicompartment electrolyzer in which the affinity gel media were packed in one of the central compartments. The presence of an electric field accelerated the migration of proteins inside the gel matrix during adsorption and desorption processes. This led to the increase of the overall speed of separation. The present study was focused on the effect of the strength of the electric field on adsorption and desorption processes.展开更多
A new and an inexpensive adsorbent of chitosan coated silica for immobilized metal affinity chromatography (IMIC) was studied. After a double coating, the chitosan coated on silica beads could be up to 53. 4 mg/g sili...A new and an inexpensive adsorbent of chitosan coated silica for immobilized metal affinity chromatography (IMIC) was studied. After a double coating, the chitosan coated on silica beads could be up to 53. 4 mg/g silica beads.When pH＞3. 8, the metal ligand Cu2+ was chelated on the coated chitosan witha bound capacity of 14. 6 mg/g chitosan without introducing iminodiacetic acid(IDA).展开更多
Absract A lipase gene, 1ip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80kDa. Lip12...Absract A lipase gene, 1ip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80kDa. Lip1233 was grouped into the lipase family X because it contained a highly conserved motif GHSLG. The recombinant enzyme was purified with Ni-NTA affinity chro- matography. The optimal temperature and pH value of Lip1233 were 45 ℃ and 8.0, respectively. It retained more than 70% of origi- nal activity after being incubated in pH ranging from 6.0 to 9.5 for 30min. It was stable when the temperature was below 45℃, but was unstable when the temperature was above 55℃. Most metal ions tested had no significant effect on the activity of Lip1233. Lip1233 remained more than original activity in some organic solvents at the concentration of 30% （v/v）. It retained more than 30% activity after incubated in pure organic solvents for 12 h, while in hexane the activity was nearly 100%. Additionally, Lip 1233 exhib- ited typical halotolerant characteristic as it was active under 4M NaC1. Lip1233 powder could catalyze efficiently the synthesis of fructose esters in hexane at 400C. These characteristics demonstrated that Lip1233 is applicable to elaborate food processing and organic synthesis.展开更多
β2-Adrenoceptor (β2-AR) was purified from the rabbit lung tissue by sepharose-salbutamol affinity chromatographic column. To prepare the β2-AR stationary phase, β2-AR was evenly immobilized on the surface of macro...β2-Adrenoceptor (β2-AR) was purified from the rabbit lung tissue by sepharose-salbutamol affinity chromatographic column. To prepare the β2-AR stationary phase, β2-AR was evenly immobilized on the surface of macro-pore silica with a mild chemical coupling method through covalent bond. The reten- tion properties of β2-AR stationary phase were characterized by four ligands, salbutamol sulfate, noradrenaline bitartrate, adrenaline hydrochloride and propranolol hydrochloride, to establish the β2-AR affinity chromatography. Then, the method was used to screen the active compounds from the total extracts of Semen Armeniacae Amarum. The results showed that β2-AR on the surface of the sta- tionary phase could keep its original bioactivity and selectivity. Amygdalin retained in the chroma- tographic column was proved to be the active compound of the total extracts of Semen Armeniacae Amarum. Compared with the existing chromatographic screening approaches, this method showed a good stability and high selectivity. The active compounds which could interact with β2-AR in traditional Chinese medicine (TCM) could be screened efficiently by this method, providing a new way to screen the active compounds in complicated samples such as TCM.展开更多
The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-...The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-β) was chosen as sense peptide and its three antisense peptides (AS-IFN 1, AS-IFN 2, and AS-IFN 3) were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide (AS-IFN 3) showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis (QCM-FIA) system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10-4, 1.31×10-4 and 2.22× 10-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.展开更多
基金grant from the Project of the Bureau of Science and Technology of Nantong (S30033).
文摘Objective： To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein （HS- AFP） for diagnosis of hepatocellular carcinoma （HCC）. Methods： HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin （LCA）-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay （RIA）. Results： Circulating AFP was separated into three peaks （AFP-1, AFP-2, and AFP-3） by LCA-affinity chromatography. Dunng the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HSAFP （AFP-3） from sera of HCC patients was eluted clearly on the LCA-sepharose gel mini-column with a solution containing α-methyI-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity （92.7%） and specificity （88.2%）. Conclusion： The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.
基金Supported by the State Key Projects(No.96c-03-04-05).
文摘A new kind of electrophoretic affinity chromatography (EAC) for bioseparation was proposed. Separation by EAC was conducted in a multicompartment electrolyzer in which the affinity gel media were packed in one of the central compartments. The presence of an electric field accelerated the migration of proteins inside the gel matrix during adsorption and desorption processes. This led to the increase of the overall speed of separation. The present study was focused on the effect of the strength of the electric field on adsorption and desorption processes.
文摘A new and an inexpensive adsorbent of chitosan coated silica for immobilized metal affinity chromatography (IMIC) was studied. After a double coating, the chitosan coated on silica beads could be up to 53. 4 mg/g silica beads.When pH＞3. 8, the metal ligand Cu2+ was chelated on the coated chitosan witha bound capacity of 14. 6 mg/g chitosan without introducing iminodiacetic acid(IDA).
基金supported by the Administration of Ocean and Fisheries of Guangdong Province (GD2012D01-002)the ‘Strategic Priority Research Program’ of the Chinese Academy of Sciences (No.XDA10030400)the Natural Science Foundation of Guangdong Province, China (Grant Nos.2015A030310270 and 2016A 030313157)
文摘Absract A lipase gene, 1ip1233, isolated from Pseudoalteromonas lipolytica SCSIO 04301, was cloned and expressed in E. coli. The enzyme comprised 810 amino acid residues with a deduced molecular weight of 80kDa. Lip1233 was grouped into the lipase family X because it contained a highly conserved motif GHSLG. The recombinant enzyme was purified with Ni-NTA affinity chro- matography. The optimal temperature and pH value of Lip1233 were 45 ℃ and 8.0, respectively. It retained more than 70% of origi- nal activity after being incubated in pH ranging from 6.0 to 9.5 for 30min. It was stable when the temperature was below 45℃, but was unstable when the temperature was above 55℃. Most metal ions tested had no significant effect on the activity of Lip1233. Lip1233 remained more than original activity in some organic solvents at the concentration of 30% （v/v）. It retained more than 30% activity after incubated in pure organic solvents for 12 h, while in hexane the activity was nearly 100%. Additionally, Lip 1233 exhib- ited typical halotolerant characteristic as it was active under 4M NaC1. Lip1233 powder could catalyze efficiently the synthesis of fructose esters in hexane at 400C. These characteristics demonstrated that Lip1233 is applicable to elaborate food processing and organic synthesis.
基金the National Natural Science Foundation of China (Grant No. 20575052)the Natural Science Foundation of Shaanxi Province, China (Grant Nos. 2006B03 and 2006C220)
文摘β2-Adrenoceptor (β2-AR) was purified from the rabbit lung tissue by sepharose-salbutamol affinity chromatographic column. To prepare the β2-AR stationary phase, β2-AR was evenly immobilized on the surface of macro-pore silica with a mild chemical coupling method through covalent bond. The reten- tion properties of β2-AR stationary phase were characterized by four ligands, salbutamol sulfate, noradrenaline bitartrate, adrenaline hydrochloride and propranolol hydrochloride, to establish the β2-AR affinity chromatography. Then, the method was used to screen the active compounds from the total extracts of Semen Armeniacae Amarum. The results showed that β2-AR on the surface of the sta- tionary phase could keep its original bioactivity and selectivity. Amygdalin retained in the chroma- tographic column was proved to be the active compound of the total extracts of Semen Armeniacae Amarum. Compared with the existing chromatographic screening approaches, this method showed a good stability and high selectivity. The active compounds which could interact with β2-AR in traditional Chinese medicine (TCM) could be screened efficiently by this method, providing a new way to screen the active compounds in complicated samples such as TCM.
基金Supported by the National Natural Science Foundation of China (Grant Nos. 20575072, 20435030 and 90408018)the Chinese Academy of Sciences (Grant No. KJCX2-SW-H06)
文摘The specific interaction between sense and antisense peptides was studied by high-performance affinity chromatography (HPAC) and quartz crystal microbalance (QCM) biosensor. Fragment 1-14 of human interferon-β (hIFN-β) was chosen as sense peptide and its three antisense peptides (AS-IFN 1, AS-IFN 2, and AS-IFN 3) were designed according to the degeneracy of genetic codes. The affinity column was prepared with sense peptide as ligand and the affinity chromatographic behavior was evaluated. Glu-substituted antisense peptide (AS-IFN 3) showed the strongest binding to immobilized sense peptide at pH 7.5. A quartz crystal microbalance-flow injection analysis (QCM-FIA) system was introduced to investigate the recognition process in real-time. The equilibrium dissociation constants between sense peptide and AS-IFN 1, AS-IFN 2 and AS-IFN 3 measured 2.08×10-4, 1.31×10-4 and 2.22× 10-5 mol/L, respectively. The mechanism study indicated that the specific recognition between sense peptide and AS-IFN 3 was due to sequence-dependent and multi-modal affinity interaction.
