• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    A serum albumin fraction is prepared in high yield and purity from plasma and other albumin-containing blood protein fractions by contacting with a resinous polymeric material having a high capacity for adsorption of albumin, heating at 65 DEG C-72 DEG C for about 1-4 hours while maintaining a pH of 5.0-5.5, and then selectively eluting the albumin from the resin-protein mixture at a pH of 3.5-4.5. The resinous polymeric materials are water-insoluble, cross-linked polyelectrolyte copolymers of ethylene and maleic anhydride containing pendant dimethylaminopropyl functional groups.
    公开号:SU736861A3
    申请号:SU772528199
    申请日:1977-07-21
    公开日:1980-05-25
    发明作者:Левис Ир Джарлс；Михаел Шук Джэмс
    申请人:Монсанто Компани (Фирма)；
    IPC主号:
    专利说明:

    EAMK copolymer. The polyelectropite copolymer is preferably converted to a hydrochloride salt to achieve greater processability of the product,
    P15 in the preferred polyelectrolyte copolymer contains about 5 methylaminobipropylamine cross-linking groups and about 90 additional dimethylaminopropylamine functional groups per 100 units. maleic anhydride in an EAMA copolymer. The polyelectrolyte copolymer is mainly capable of fully adsorbing the blood fraction albumin, and then the albumin can be isolated with a yield higher than 90% with 94% purity using the method of hot treatment and selective elution.
    The polyelectrolyte copolymers are mixed with blood plasma or serum, or albumin-containing blood fractions, preferably at concentrations of 1-5% copolymer. By adjusting the pH of the resinous protein mixture to different levels, selective proteins are first removed. At pH 5.5-7.5, albucine, L and jS-globulins and fibrinogen are adsorbed by the resin, while a large part of the U -globulin remains unabsorbed and can be isolated from the residue for therapeutic purposes. The initial release of jp -globulin is carried out at pH 6. Part of the absorbed ir-globulins and fibrinogen are recovered by adjusting the pH of the resinous protein mixture to about 4.7, followed by collecting the desorbed residue.
    The pH of the resinous protein mixture is then adjusted to 5.0-5.5 with heating to a temperature of about 65-72 0 for 1-4 hours, the pH is adjusted to 5.2-5.3 with heating of the resinous protein mixture at about 72 ° C for 1 hour
    In the hot processing stage, residual globulins, mainly (/ h-globulins, are denatured, while albumin is not denatured and is easy to release. After the heat treatment, the pH is adjusted to 3.5-4.5 for elution from the resin the albumin mixture of the desired albumin. The resinous protein mixture is cooled before pH adjustment, the pH value is adjusted to 4, gchumin is isolated by precipitation, filtration or centrifugation, the preferred method is a resinous protein mixture filter nym pH, flushing and collecting the filtrate as the desired albumin coat:. tion, high purity Adjustment of pH to the required level is carried out by treatment with an acid
    or alkaline buffer, e.g. sodium acetate acetate or citric acid based buffer, for acidification purposes or by treatment with sodium bicarbonate or sodium hydroxide for alkalization. Preferred is the use of albumin stabilizers, such as sodium acetyl tryptophanate and sodium caprylate resin-protein mixture.
    Q treatment time heating with regard to their stabilizing properties.
    Example 1 A polyelectrolyte polymer consists of a resinous product of reacting mostly equimolar parts of ethylene with anhyd5 REDOM maleic acid (AMK), transversely methylimino-propropylamine (MIBPA) and then subjected to interaction with dimethylaminopropylamine (DMPA) in this way
    0 to form about, five mibpashivayuschih groups and about 90 DMPA additional groups per 100 units. AMC in LMK copolymer, and converted into hydrochloric acid salt. First, the polyelectrolytic rolite copolymer is washed in
    0.04 M sodium chloride. Plasma obtained from mixed human blood is diluted with 3 parts of water per 1 part of plasma, after which 2% by weight of the washed poly-electrolyte copolymer is mixed (2 g / 100 ml). The pH of the resin-plasma mixture is adjusted to 6.0, then stirred for 30 minutes, filtered and
    washed with 0.002 M sodium chloride. The filtrate, mainly consisting of y-globulins, p -globulins, fibrinogen and associated blood factors, is isolated from the obtained resinous protein residue on the filter,
     The latter, containing the adsorbed albumin, is acidified to a pH of 5.2. To achieve a 0.012 M concentration, a proper amount of sodium caprylate-based stabilizer is admixed to the adsorbed RESIN protein, sodium chloride is added to achieve the 0.002 M concentration. After that, for 1 h at 10 ° C, the protein absorbed by the resin is heated, then the material is filtered and the filtrate is discarded.
    Albumin is eluted from the remaining gummy protein by acidification to pH 4.0 in 0.002 M sodium chloride with citric acid, then peremanyat and filtered. The filtrate is collected as the desired albumin fraction with a yield of 96.6% (based on the concentration of albumin in the initial plasma) with a purity of 98.5%. The degree of purity of albumin eluted from the resin is determined by agar-gel electrophoresis in a buffer
    5 on the basis of dystilbarbiturovuyu acid at pH 8.6 using an electrophoretic device. The counting is performed at 600 nm using a densitometer. Blue Soiahayche BrAttiAnt R 250 is a protein sensitivity agent of high sensitivity. Due to the high degree of sensitivity, the blue type Coomasste Br- «e, BEue K 250 allows the determination of protein npfiMeceft in the albumin product with very high accuracy, and the quantitative analysis performed confirms the production of albuminia with a high degree of purity.
    Example 2. The method of Example 1 is repeated, except that the starting plasma is an AHF-depleted plasma. The yield and purity of the albumin product are basically the same as in Example 1.
    Example 3. The method of Example 1 is repeated, except that I includes an intermediate stage between the first filtration and hot treatment in order to remove additional globulins and fibrinogen from the plasma. At this intermediate stage, the pH of the resinous protein residue on the filter from the first filtration is adjusted to 4.7 with 0.002 M sodium chloride citric acid, then stirring for 30 minutes. The mixture is filtered, progressed and then subjected to heat treatment, then to the steps of Example 1. The filtrate from the treatment with a pH of 4.7 contains A and -globulins and fibrinogen, which are collected for use in various known therapeutic and diagnostic purposes. Target albumin is obtained with a yield and degree of purity generally similar to Example 1.
    Following the release of the purified .ot-o albumin product, the albumin is concentrated to the desired level, the pH of the cocoa-concentrated product is adjusted to a physiologically acceptable value and the electrolyte content, then heated to destroy the viruses, brought to a transparent state by filtration or other means to obtain a clinically a direct substance complying with the requirements for a usually whey material. For the preferred method of concurrent esh used in practice, it is based on lyophiosis and followed by fading to the desired end of the walkie-talkie, for example, 5 5 or 25% w.rfrafiltration.
    Offering 1st with fso.b allows you to increase the yield of the target product with a value of 96.6% at a degree of 98.5%.
    20
    权利要求:
    Claims (1)
    [1]
    1. US Patent 3555001, cl. 266-112, pub. 1969.
    类似技术:
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    同族专利:
    公开号 | 公开日
    SE7708406L|1978-01-23|
    NL7708005A|1978-01-24|
    MX4620E|1982-07-21|
    CA1069820A|1980-01-15|
    HU175972B|1980-11-28|
    DE2732998A1|1978-01-26|
    RO72952A|1982-05-10|
    GB1543111A|1979-03-28|
    FR2358886B1|1981-10-30|
    US4097473A|1978-06-27|
    JPS5315414A|1978-02-13|
    AT362068B|1981-04-27|
    AU2720177A|1979-01-25|
    BE856991A|1978-01-20|
    JPS6034521B2|1985-08-09|
    AU513473B2|1980-12-04|
    DE2732998C2|1986-12-04|
    SE442589B|1986-01-20|
    CH633019A5|1982-11-15|
    ATA528677A|1980-09-15|
    FR2358886A1|1978-02-17|
    IT1081299B|1985-05-16|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    US2469193A|1942-02-09|1949-05-03|Research Corp|Protein fractionation|
    US2705230A|1949-09-23|1955-03-29|Allen F Reid|Method of purifying albumin|
    US2765299A|1952-06-27|1956-10-02|Armour & Co|Recovery of serum albumin|
    US2958628A|1957-02-21|1960-11-01|Cutter Lab|Heat treatable plasma protein product and method of preparation|
    US3100737A|1958-02-05|1963-08-13|Auerswald Wilhelm|Method of preparing a plasma protein solution free of active hepatitis virus and product produced thereby|
    US3554985A|1963-01-02|1971-01-12|Monsanto Co|Cross-linked copolymer polyelectrolytes based on alpha,beta-ethylenically unsaturated acids|
    US3555001A|1969-05-29|1971-01-12|Monsanto Co|Process for the fractionation of plasma and serum using water-insoluble polyelectrolytes containing diloweralkylaminoloweralkylimide groups|JPS5843369B2|1977-09-10|1983-09-27|Hidetoshi Tsuchida|
    US4305871A|1980-09-02|1981-12-15|Edward Shanbrom|Method of selectively increasing yield and purity of certain cryoprecipitate proteins by heating|
    WO1983002057A1|1981-12-17|1983-06-23|Quinn, Patrick, James|Contraceptive method and composition|
    US4397841A|1982-06-28|1983-08-09|Monsanto Company|Production of blood coagulation factor VIII:C|
    US4382028A|1982-07-19|1983-05-03|Monsanto Company|Separation of plasma proteins from cell culture systems|
    HUT40311A|1983-06-03|1986-12-28|Kiskunhalasi Aag|Process for producing protein concentrates, blood-curd and nutriments from blood and its elements|
    US4533496A|1984-05-08|1985-08-06|Monsanto Company|Method of isolating monoclonal antibodies from hybridoma cultures|
    GB8628104D0|1986-11-25|1986-12-31|Connaught Lab|Pasteurization of immunoglobin solutions|
    US5277818A|1988-10-31|1994-01-11|The Green Cross Corporation|Albumin preparation and process for preparing the same|
    US5250662A|1989-10-05|1993-10-05|Alpha Therapeutic Corporation|Albumin purification|
    FR2686620B1|1992-01-27|1995-06-23|Rhone Poulenc Rorer Sa|HUMAN SERUM-ALBUMIN, PREPARATION AND USE.|
    KR20120018827A|2009-02-20|2012-03-05|벤트리아 바이오사이언스|Cell culture media containing combinations of proteins|
    法律状态:
    优先权:
    申请号 | 申请日 | 专利标题
    US70790676A| true| 1976-07-22|1976-07-22|
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