US3706660A - Process for removal of particle-forming proteins from blood sera - Google Patents
Process for removal of particle-forming proteins from blood sera Download PDFInfo
- Publication number
- US3706660A US3706660A US128152A US3706660DA US3706660A US 3706660 A US3706660 A US 3706660A US 128152 A US128152 A US 128152A US 3706660D A US3706660D A US 3706660DA US 3706660 A US3706660 A US 3706660A
- Authority
- US
- United States
- Prior art keywords
- serum
- accordance
- straight chain
- chain hydrocarbon
- particle
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/06—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from blood
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
- G01N33/491—Blood by separating the blood components
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/827—Proteins from mammals or birds
- Y10S530/829—Blood
- Y10S530/83—Plasma; serum
Definitions
- Blood sera for grouping and typing purposes must be pure and substantially free of particle-forming substances. Formation of proteinaceous particles in blood serum, withor without fibrinogen, occurs when'the serum is shaken, for example, for 2 to 5 hours.
- Several methods have been suggested for removing particleforming proteins from sera.
- British Pat. No. 1,196,636 discloses a process for preparing pure serum albumins from normal and hemolyzed sera and plasma by precipitating undesirable proteins and dyestuffs contained in a solution of raw albumins by heating a 5-8 percent solution of raw albumin together with percent by volume of chloroform at a pH of 4.95-5.05 to a temperatureof 50 C.
- serum albumin of standard quality can be obtained by a method of ethanol fractionation as described by Cohn et al., J. Am. Chem. Soc., 68, 3386, (1946)and 72,465,(l950).
- a process for substantially preventing or retarding the formation of proteinaceous particles in blood serum induced by shaking, by removal of particle-forming proteins therefrom, which comprises adjusting pH of the serum to within the range of from about 5.2 to about 5.6 and preferably from about 5.2 to about 5.4, mixing the blood serum with a halogenated straight chain hydrocarbon, thereby causing the formation of a proteinaceous precipitate, and separating the precipitate from the serum.
- the halogenated straight chain hydrocarbon in employed in a volume ratio to the serum of within the range of from about 1:2 to about 2:1 or greater; however, no apparent benefit is obtained employing ratios greater than 5:1.
- the pH of the serum can be adjusted to the required level before or after the addition of the halogenated straight chain hydrocarbon thereto by admixing with dilute acid such as a mineral acid, for example hydrochloric acid, sulfuric acid or nitric acid, or an organic acid such as formic acid or acetic acid.
- dilute acid such as a mineral acid, for example hydrochloric acid, sulfuric acid or nitric acid, or an organic acid such as formic acid or acetic acid.
- the blood serum once adjusted to a pH within the range of from about 5.2 to about 5.6, can be shaken to induce precipitation of proteinaceous particles prior to adding the halogenated straight chain hydrocarbon thereto.
- the extraction is usually carried out at ambient temperature, although, if desired, temperatures ranging from about to about 35 C can be employed to expedite formation of the proteinaceous precipitate.
- the mixture Upon mixing of the hydrocarbon solvent with the serum having a pH within the above defined range the mixture is shaken or mixed for a period ranging from about A to about 1 k hours to ensure complete admixture. Longer mixing periods can be employed, with no apparent benefit.
- the mixture After mixing, the mixture is allowed to settle preferably at reduced temperatures ranging from about 0 to about 20 C, for periods ranging from about 1 to about 20 hours.
- the precipitate formed in the mixture can then be separated, for example, by centrifugation or other conventional technique.
- the clear serum layer can then be removed and further purified, for example, by placing the serum under vacuum at a temperature within the range of from about 20 to about 40 C with stirring until foaming ceases, adjusting the pH of serum to within the range of from about 6.5 to about 8, for example by adding alkali thereto, such as sodium hydroxide, potassium hydroxide, sodium carbonate, ammonium hydroxide or tris(hydroxymethyl)aminomethane.
- the serum can then be treated with an absorbent such as'kaolin, further centrifuged and filtered to further purify the serum.
- halogenated straight chain hydrocarbon solvents which can be employed herein include chloroform, methylene chloride or carbon tetrachloride.
- EXAMPLE 1 The pH or human serum (plasma with substantially all fibrinogen removed therefrom) is adjusted to a value of 5.2 by adding about 1.2 ml of dilute acetic acid (30 percent), to ml of serum. An equal volume of methylene chloride is added to the serum and the mixture is shaken for three hours at 30 C. The mixture is allowed to stand at room temperature for several hours and then is centrifuged at 5,000 X g for 10 minutes whereupon three layers form, a solvent layer, a layer containing a solid precipitate and a serum liquid layer. The three layers are separated and the serum layer is placed under vacuum at 35 with stirring (for at least 15 minutes) until foaming ceases. The pH of the serum is adjusted. to 7.2 by adding 5N sodium hydroxide thereto. The serum is then filtered through a sterile D-8 filter pad to thereby remove additional impurities therefrom.
- EXAMPLE 2 The pH of human serum (plasma with substantially all fibrinogen removed therefrom) is adjusted to a value of 5.5 by adding about 0.9 ml of dilute acetic acid (30 percent), to 100 ml of serum. An equal volume of chloroform is added to the serum and the mixture is shaken for 5 hours at 45 C. The mixture is allowed to stand at room temperature for several hours and then is centrifuged at 10,000 X g for 10 minutes whereupon three layers form, a solvent layer, a layer containing a solid precipitate and a serum liquid layer. The three layers are separated and the serum layer is placed under vacuum at 35 with stirring (for at least 15 minutes) until foaming ceases. The pH of the serum is adjusted to 6.5-8 by adding 5N sodium hydroxide thereto. The serum is then filtered through a sterile D-8 filter pad to thereby removed additional impurities therefrom.
- EXAMPLE 3 The pH of human serum (plasma with substantially all fibrinogen removed therefrom) is adjusted to a value of 5.4 by adding about 1 ml of dilute acetic acid (30 percent), to 100 ml of serum. About 200 ml of chloroform is added to .the serum and the mixture is shaken for 3 hours at 25 C. The mixture is allowed to stand at room temperature for several hours and then is centrifuged at 7,000 X g for 10 minutes whereupon three layers form, a solvent layer, a layer containing a solid precipitate and a serum liquid layer. The three layers are separated and the serum layer is placed under vacuum at 35 with stirring (for at least 15' minutes) until foaming ceases.
- the pH of the serum is adjusted to 7.2 by adding 5N sodium hydroxide thereto.
- About 1 g kaolin is added to the serum and the mixture isstirred on a magnetic stirrer for 1% hours.
- the serum is then filtered through a sterile D-8 filterer pad to thereby remove additional impurities therefrom.
- EXAMPLE 4 The pH of human serum (plasma with substantially all fibrinogen removed therefrom) is adjusted to a value of 5.4 by adding about 1 ml of dilute acetic acid (30 percent), to 100 ml of serum. The serum is shaken for 3 hours at a temperature of about 35 C. and the resulting precipitate removed. About 300 ml of chloroform is mixed with the supernatent and the mixture is allowed to stand at room temperature for several hours and then is centrifuged at 7,000 X g for 10 minutes whereupon three layers form, a solvent layer, a layer containing a solid precipitate and a serum liquid layer. The three layers are separated and the serum layer is placed under vacuum at 35 with stirring (for at least minutes) until foaming ceases.
- the pH is adjusted to' 7.2 by adding 5N sodium hydroxide thereto.
- About 1 g kaolin is added to the serum and the mixture is stirred on a magnetic stirrer for 1% hours.
- the serum is then filtered through a sterile D-8 filter pad to thereby remove additional impurities therefrom.
- the treated human serum obtained in each of examples l to 4 is substantially free of particle-forming protein and is acceptable for grouping and Rh typing purpose. Furthermore, the antibody titre of the serum is not significantly affected by the procedure described in these examples.
- a process for substantially preventing or retarding the formation of particles in blood sera induced by shaking, by removal of particle-forming proteins therefrom which comprises adjusting pH of the blood serum to within the range of from about 5.2 to about 5.6, mixing or extracting the blood serum at ambient temperature up to about 35 C with from about 0.5 to about 5 volumes of a halogenated straight chain hydrocarbon per volume of blood serum, thereby causing the formation of a proteinaceous precipitate, and separating the precipitate from the serum.
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Food Science & Technology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Ecology (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12815271A | 1971-03-25 | 1971-03-25 |
Publications (1)
Publication Number | Publication Date |
---|---|
US3706660A true US3706660A (en) | 1972-12-19 |
Family
ID=22433894
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US128152A Expired - Lifetime US3706660A (en) | 1971-03-25 | 1971-03-25 | Process for removal of particle-forming proteins from blood sera |
Country Status (7)
Country | Link |
---|---|
US (1) | US3706660A (de) |
CA (1) | CA955851A (de) |
CH (1) | CH551011A (de) |
DE (1) | DE2214260A1 (de) |
FR (1) | FR2131635A5 (de) |
GB (1) | GB1353847A (de) |
HU (1) | HU165141B (de) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3868220A (en) * | 1973-08-13 | 1975-02-25 | Squibb & Sons Inc | Process for extracting procainamide from blood |
US3958939A (en) * | 1975-01-08 | 1976-05-25 | Coulter Electronics, Inc. | Method for clarification of lipemic serum |
EP0456326A1 (de) * | 1990-05-07 | 1991-11-13 | Harimex-Ligos B.V. | Verfahren zur Reinigung von Blutplasma |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4264471A (en) * | 1979-06-04 | 1981-04-28 | E. I. Du Pont De Nemours And Company | Serum and plasma clarification process |
DE3344656A1 (de) * | 1983-12-09 | 1985-06-13 | Lentia GmbH Chem. u. pharm. Erzeugnisse - Industriebedarf, 8000 München | Verfahren zur herstellung einer serumproteinloesung |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1196636A (en) * | 1967-03-01 | 1970-07-01 | Ustav Ser A Ockovacich Latek O | Process for the Preparation of Pure Albumins from Normal and Haemolysed Sera and Plasma |
-
1971
- 1971-03-25 US US128152A patent/US3706660A/en not_active Expired - Lifetime
-
1972
- 1972-03-13 CA CA136,933A patent/CA955851A/en not_active Expired
- 1972-03-14 GB GB1190772A patent/GB1353847A/en not_active Expired
- 1972-03-15 CH CH382572A patent/CH551011A/fr not_active IP Right Cessation
- 1972-03-23 DE DE19722214260 patent/DE2214260A1/de active Pending
- 1972-03-24 FR FR7210562A patent/FR2131635A5/fr not_active Expired
- 1972-03-24 HU HUSU727A patent/HU165141B/hu unknown
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB1196636A (en) * | 1967-03-01 | 1970-07-01 | Ustav Ser A Ockovacich Latek O | Process for the Preparation of Pure Albumins from Normal and Haemolysed Sera and Plasma |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3868220A (en) * | 1973-08-13 | 1975-02-25 | Squibb & Sons Inc | Process for extracting procainamide from blood |
US3958939A (en) * | 1975-01-08 | 1976-05-25 | Coulter Electronics, Inc. | Method for clarification of lipemic serum |
EP0456326A1 (de) * | 1990-05-07 | 1991-11-13 | Harimex-Ligos B.V. | Verfahren zur Reinigung von Blutplasma |
US5252221A (en) * | 1990-05-07 | 1993-10-12 | Harimex-Ligos B.V. | Method for purifying blood plasma |
Also Published As
Publication number | Publication date |
---|---|
CH551011A (fr) | 1974-06-28 |
DE2214260A1 (de) | 1972-10-26 |
FR2131635A5 (de) | 1972-11-10 |
HU165141B (de) | 1974-06-28 |
CA955851A (en) | 1974-10-08 |
GB1353847A (en) | 1974-05-22 |
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