IgE is one of the five immunoglobulin classes in human serum, with molecular weigh about 190 kD. The range of IgE in atopic patient is at least 30-40 folds more than normal population. In addition to difficulties of PURIFICATION, the expense is very high; so we attempted to use a relative simple and economic method for PURIFICATION of IgE.In present study, high titer patients serum (higher than 1000 IU/ml) were collected and pooled. Serum diluted with phosphate buffer saline (PBS) and precipitated with 40% ammonium sulfate. Then, the IgE rich supernatant were precipitated with 60% ammonium sulfate. The precipitate dialysed against phosphate buffer and run on the ion exchange column. IgE rich fractions were eluted with NaCl gradient. To remove any contaminant proteins, IgE rich fractions were run on affinity column with sepharose 4B anti-IgE antibody. The absorbed IgE was eluted with changing of buffer. Finally, the purified IgE approved with SDS-PAGE and immunobloting methods.

Solvent extraction is an economically efficient method widely used in the PURIFICATION of wet phosphoric acid. In this study, a microchannel was applied to promote the mixing and PURIFICATION of phosphoric acid during continuous production. For this aim, solvent extraction was conducted to purify phosphoric acid via methyl isobutyl ketone/tri-butyl phosphate mixtures. Additionally, the Box-Behnken design method was used to survey the solvent extraction process. The effect of various operational parameters such as solvent concentration (45– 65 %wt. ), temperature (18-28˚ C), and organic/aqueous phase ratio (2: 1– 6: 1) at a constant flow rate of 70 mL/L was examined. Experimental results indicated that the microchannel at the residence time of 6. 85 min could promote the extraction percentage of and sulfate removal of more than 98% and 60%, respectively, compared to the batch extractor.

Introduction: Peroxides is a glycoprotein that contains hem group and variable cationic and anionic is enzymes. This enzyme is used in laboratory diagnosis for determination of hormones and bacterial toxins with ELISA method, detection of tissue antigens and macrophage activation against tumoral cells. In addition, this enzyme is used for measurement of glucose, urea, uric acid and cholesterol. Although peroxides is widely seen in available sources of plants, this enzyme for meeting the consuming needs of our country is imported from abroad with high foreign currency costs.Objective: This experimental study was conducted for separation and PURIFICATION of peroxides is enzymes from cultivated radish which is cheap and abundant in all seasons in Iran.Materials and Methods: Extraction was carried out by homogenization of plant roots and concentration of Ammonium sulfate. Then, PURIFICATION method was followed by gel filtration, cationic exchange chromatography. Finally, separation of basic and acidic is enzymes were conducted by different and repeated column chromatography and SDS-PAGE.Results: As a result of these procedures, PURIFICATION degree obtained was 168.5 folds more than the raw extract and specific activity was comparable with commercial peroxides.Conclusion: As this enzyme is stable at wide range of PH and temperature, we can economize its PURIFICATION methods to a great extent and achieve the maximum required foreign currency savings. It is hoped that with the use of wide and cheap sources, we can take an effective step toward large scale production of the enzyme in our country and avoid spending our financial resources in purchasing peroxides from foreign countries.

Background and Objective : The genus of sarcocystis, zonotic parasites, have two hosts in their life cycle. They have also special importance in industrial veterinary. The serological tests are the best methods for detection of the parasite. This research was planned for isolation of sheep sarcocystis specific protein for using in serological laboratory tests. Materials and Methods : The infected muscles of sheep carcass were collected from Tehran slaughter house and transferred to our laboratory. The sarcocystis were isolated from the infected muscles and crude antigen was prepared . The crude antigen was fragmented by serial dilution of ammonium sulfate solution and followed by size exclusion chromatography. Results: Crude antigen was electrophoresed by SDS-PAGE and its protein bands were detected by commassi brilliant blue staining. We used different concentrations of ammonium sulfate for precipitation and after by size exclusion chromatography, a 35kDa protein band was separated and observed by SDS-PAGE. Conclusion: The protein band of sheep sarcocystis which can be used as antigen in serological methods was parified and detected.

Insulin is a two-chain polypeptide hormone with molecular weight of 5.7 KD, which has 51 AA. By some disulfide bounds, these two chains are connected to each other. One of the most important diseases associating insulin is diabetes. In these patients, in the absence of insulin, cells cannot absorb glucose and glucose level of blood will increase and finally body tissues encounter a great energy shortage causing different acute and chronic implications. Nowadays for measuring this hormone, RIA methods are used. The aim of this research is designing of ELISA kit for measuring of insulin and the first step in designing such kits consists of providing and purifying antibody against human insulin. In order to provide and purify polyclonal antibody to insulin some practical fulfilled as follows. For produce antibody, we immunized Guinea pigs and after immunization stages, they were bleeded and their serum was separated. In order to assess and diagnose the produce anti-insulin by these pigs, RIA, DID and Dot blot methods were used. After making sure of the presence of insulin antibody in the serum, we purified antibody with saturated ammonium sulfate and then ion exchange chromatography. Also for the assessment of the antibody PURIFICATION rate, the SDS-PAGE technique was used. Finally Western Blotting technique was used to make sure that there is not any unspecified bound.

Abstract‎ Objective: Aim of this study was to purify Human serum paraoxonase-1, which is one of the ‎important organophosphate detoxifying enzymes. Materials and Methods: In this study we found anion exchange DEAE Sephadex A-50 and gel ‎filtration media Sephadex G-200 is suitable for PURIFICATION of this enzyme. Using Triton X-100 ‎a non ionic detergent, enzyme was separated from lipoproteins. Then enzyme solution was ‎applied to DEAE column and washed to elute non specific binding. The bind enzyme was eluted ‎with sodium chloride gradient. Active fractions were pooled and applied to G-200 column. The ‎active fractions were applied to the second DEAE column. Changing the buffer and the pH ‎resulted to a purified enzyme. Results: Results of these chromatographic procedures showed purified enzyme with greater than ‎‎95% purity and 320 U/L of specific activity. SDS-Poly acryl amid gel electrophoresis showed a ‎single band of approximately 43 KD protein. Kinetic properties of the purified enzyme showed ‎that, affinity of the enzyme to the substrates is dependent on buffer composition and the calcium ‎and sodium ions concentration. Using paraoxon as the substrate the activity was related to the ‎calcium and sodium concentration (Km=1.39±0.52), but for phenyl acetate as the substrate the ‎activity was independent of sodium and calcium (Km=0.728±0.105). The optimal pH for ‎paraoxon hydrolysis was around 9.5-11, but for phenyl acetate it was from 8-8.5. Glycerol 20% ‎‎(v/v) was most effective stabilizer. Keeping at 25oC for 20 days 75% of the original activity was ‎restored. Conversely at 56oC the ammonium sulfate was better stabilizer. EDTA and zinc ‎chloride both inhibited the enzyme activity. Concomitant incubation of the inhibitors with ‎calcium ion resulted to increased IC50 values.‎ Conclusion: The stablished PURIFICATION procedure is simple and non-expensive and can be used ‎for large scale PURIFICATION of this enzyme which can be used for organophosphate ‎decontamination or prevention of intoxication.

Although many environmental problems such as pollution shortage of natural resources and damages to the environment are apparently results of mans unplanned and wrong use of nature the root of the problems should be sought from neglecting the spirituality and (symbolic) meaning of nature and its elements which have been considered in all traditional cultures. The quantitative thought predominated over the modern era (like modern architecture which considers buildings and cities as machines) approaches the nature as a machine from which the maximum economic and material efficiency is the final aim.Researching the hypothesis which considers effects of promotion and revitalization of spiritual culture on protecting environment and elements deals with three main components. "culture" "natural elements" and "geographical place". In this research "Iranian culture" "water" and "Iran" are respectively selected as justifications of the above mentioned components. The mechanical school of thought takes into water as a part of the machine of existence by which human physiological and material needs can be responded. Whereas by paying attention to different degrees of human life (spiritual mental and material) and by considering the importance of spiritual and mental aspects of human life everything in the world is meaningful. Being away of the fact that nature and natural elements have (symbolic and spiritual) meanings will help straighten and enrich in addition to being a guide in the quantitative field in relation to nature. Basic subjects in Iranian culture will be considered in this paper. Moreover the paper endeavors to investigate the ways of getting benefits from historical experiments by which besides solving material problems human life will be meaningful. The main titles (subjects) of the paper are as follows: "introduction and generalities" "spiritual aspects of water in Iranian culture" "water in Iranian architectural and urban spaces" "water and its related architectural elements" "present situation" and "conclusion and suggestions".

Streptokinase is an extracellular protein, extracted from certain strains of beta hemolytic streptococcus. It is a non-protease plasminogen activator that activates plasminogen to plasmin, the enzyme that degrades fibrin cloth through its specific lysine binding site; it is used therefore as a drug in thrombolytic therapy. The rate of bacterial growth and streptokinase production was studied in condition of excess glucose addition to culture media and its pH maintenance. The streptokinase product of the bacterial culture was preliminary extracted by salt precipitation and then purified by affinity chromatography on plasminogen substituted sepharose-4B in a condition that the plasminogen active site was protected from streptokinaseinduced activation. The purity of streptokinase was confirmed by SDS-PAGE and its biological activity determined in a specific streptokinase assay. The results showed that in the fed –batch culture, the rate of streptokinase production increased over two times as compared with the batch culture while at the same time, shortening the streptokinase PURIFICATION to a single step increased the yield over 95 % at the chromatography stage.

Background: Pseudomonas putida (P. putida) ATCC12633 can produce creatinase. It is a microbial enzyme which degrades creatinine in bacteria and provides source of carbon and nitrogen. Also, this enzyme is used in the enzymatic measurement of creatinine concentration for diagnosis of renal and muscles functions and diseases. Our purpose was recombinant production of creatinase for using in clinical measurement of serum or urine creatinine.Methods: A 1209bp of open reading frame of creatinase was amplified by PCR from P. putida ATCC12633genome and cloned into pET28a expression vector which was digested using NheI and XhoI restriction enzymes. Cloning was confirmed by colony PCR, double digestion analysis and sequencing. Recombinant pET28a vector was transformed toEscherichia coli (E. coli) BL21 (DE3). Creatinase expression was induced inE.coli BL21 (DE3) using IPTG and confirmed by SDS-PAGE and western blotting.PURIFICATION of creatinase was performed using Ni-NTA column. The specific activity of this enzyme was also investigated.Results: The creatinase gene cloning was confirmed by DNA sequencing. Successful expression of creatinase was performed inE. coli (57.4% of total protein). SDS-PAGE and western blot analysis showed a 45kDa creatinase protein. PURIFICATION of creatinase was done with high purity. The specific activity of recombinant enzyme is 26.54 unit/mgthat is much higher than other creatinase used in the commercial kits (9 unit/mg).Conclusion: The P. putida ATCC12633 recombinant creatinase was expressed efficiently inE. coli BL21 and 57% of total protein was the recombinant creatinase. Also, expressed creatinase has high solubility and also the enzyme has good activity compared to enzymes used in commercial kits, so a new source of creatinase was produced for creatinine assay kit in this study.