US3852415A - Compositions for use in radioimmunoassay as a substitute for blood plasma extract in determination of carcinoembryonic antigen - Google Patents
Compositions for use in radioimmunoassay as a substitute for blood plasma extract in determination of carcinoembryonic antigen Download PDFInfo
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- US3852415A US3852415A US00297565A US29756572A US3852415A US 3852415 A US3852415 A US 3852415A US 00297565 A US00297565 A US 00297565A US 29756572 A US29756572 A US 29756572A US 3852415 A US3852415 A US 3852415A
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- 239000000203 mixture Substances 0.000 title claims abstract description 41
- 210000002381 plasma Anatomy 0.000 title claims abstract description 40
- 102000012406 Carcinoembryonic Antigen Human genes 0.000 title abstract description 39
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 title abstract description 39
- 239000000284 extract Substances 0.000 title abstract description 30
- 238000003127 radioimmunoassay Methods 0.000 title description 3
- 230000005764 inhibitory process Effects 0.000 claims abstract description 16
- 238000000954 titration curve Methods 0.000 claims abstract description 16
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 27
- 239000003085 diluting agent Substances 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 239000003755 preservative agent Substances 0.000 claims description 15
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical group [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 13
- 230000002335 preservative effect Effects 0.000 claims description 13
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 11
- 229940098773 bovine serum albumin Drugs 0.000 claims description 11
- 239000000872 buffer Substances 0.000 claims description 9
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 7
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- 239000004599 antimicrobial Substances 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 6
- 239000010452 phosphate Substances 0.000 claims description 6
- 230000000845 anti-microbial effect Effects 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 229910052783 alkali metal Inorganic materials 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 4
- FXKZPKBFTQUJBA-UHFFFAOYSA-N 2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;sodium;dihydrate Chemical compound O.O.[Na].[Na].OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O FXKZPKBFTQUJBA-UHFFFAOYSA-N 0.000 claims description 3
- 150000001340 alkali metals Chemical class 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 150000001735 carboxylic acids Chemical class 0.000 claims 1
- 150000004683 dihydrates Chemical class 0.000 claims 1
- PMNYTGAGAKEGJE-UHFFFAOYSA-N ethane-1,2-diamine;sodium Chemical compound [Na].[Na].NCCN PMNYTGAGAKEGJE-UHFFFAOYSA-N 0.000 claims 1
- 239000000427 antigen Substances 0.000 abstract description 5
- 102000036639 antigens Human genes 0.000 abstract description 4
- 108091007433 antigens Proteins 0.000 abstract description 4
- 210000004369 blood Anatomy 0.000 description 11
- 239000008280 blood Substances 0.000 description 11
- 239000002585 base Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 239000008367 deionised water Substances 0.000 description 7
- 239000012153 distilled water Substances 0.000 description 7
- 239000011521 glass Substances 0.000 description 5
- 150000007524 organic acids Chemical class 0.000 description 5
- 239000003109 Disodium ethylene diamine tetraacetate Substances 0.000 description 4
- -1 alkali metal salts Chemical class 0.000 description 4
- 238000000502 dialysis Methods 0.000 description 4
- 235000019301 disodium ethylene diamine tetraacetate Nutrition 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 3
- 239000005695 Ammonium acetate Substances 0.000 description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 235000019257 ammonium acetate Nutrition 0.000 description 3
- 229940043376 ammonium acetate Drugs 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical compound [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000003058 plasma substitute Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 description 1
- DEXFNLNNUZKHNO-UHFFFAOYSA-N 6-[3-[4-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]piperidin-1-yl]-3-oxopropyl]-3H-1,3-benzoxazol-2-one Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1CCN(CC1)C(CCC1=CC2=C(NC(O2)=O)C=C1)=O DEXFNLNNUZKHNO-UHFFFAOYSA-N 0.000 description 1
- BDDLHHRCDSJVKV-UHFFFAOYSA-N 7028-40-2 Chemical class CC(O)=O.CC(O)=O.CC(O)=O.CC(O)=O BDDLHHRCDSJVKV-UHFFFAOYSA-N 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000000385 dialysis solution Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical class [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 150000007519 polyprotic acids Polymers 0.000 description 1
- 235000010241 potassium sorbate Nutrition 0.000 description 1
- 239000004302 potassium sorbate Substances 0.000 description 1
- 229940069338 potassium sorbate Drugs 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Images
Classifications
-
- 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
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard
-
- 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
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/967—Standards, controls, materials, e.g. validation studies, buffer systems
-
- 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
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/811—Test for named disease, body condition or organ function
- Y10S436/813—Cancer
-
- 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
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/106664—Blood serum or blood plasma standard or control
-
- 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
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/107497—Preparation composition [e.g., lysing or precipitation, etc.]
-
- 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
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/108331—Preservative, buffer, anticoagulant or diluent
Definitions
- FIG. 1 ANTIBODY TITRATION CURVES I l I I AA PLASMA EXTRACT I50 PLASMA EXTRACT SUBSTITUTE I30 W 1/0 cpm x IO' 90 0 50 I00 200 500 ,u! Of ANT/SERUM (l' 2500 dill?) FIG. 1
- a diluent medium a material which will permit consistent, reliable results.
- plasma extracted by glycoprotein solvents and dialysis from normal blood group Type A+ blood was used as the diluent medium.
- Such as extraction process is disclosed, for example, in US. Pat. No. 3,663,684, Example 5. While the results using this diluent are satisfactory, there is difficulty in obtaining sufficient quantities of normal plasma extract without an interfering amount of CEA.
- FIG. 1 shows antibody titration curves.
- FIG. 2 shows standard inhibition curves.
- This substitute is a buffer composition containing as the buffer, a salt of an organic acid and a strong inorganic base, a preservative, a protein to coat glass tubes, a base and water.
- the concentrations and amounts of ingredients as well as pH must be adjusted so the curve is substantially identical to that resulting when plasma extract from blood group Type A+ is used.
- the substitute plasma extract composition when used to make a standard inhibition curve must produce a curve which is substantially identical to that resulting when plasma extract from blood group Type A+ is used.
- this range includes the pKs of polybasic acids. For purposes of this invention, however, only the pK and pK are significant.
- a preferred pK is in the range of about 2 to 3.
- Other criteria for choosing a suitable organic acid are that it must form water soluble salts with alkali metals, and it must cause the pH of the composition to be near neutral on the acid side.
- Typical, suitable, organic acids are the tetraacetic acid compounds, particularly ethylenediamine tetraacetic acid (EDTA) which has a pK of 2.0 and a pK of 2.7.
- EDTA ethylenediamine tetraacetic acid
- Other similar organic acids are suitable, for example, diethylenetriamine pentacetic acid.
- Preferred alkali metal salts are the disodium and dipotassium salts.
- the concentration of the organic acid should be equivalent in function to 1.28-1.32 gm./liter of disodium EDTA. Preferably, 1.3 gm./liter is required. In the event a tenfold concentration increase of the plasma extract substitute per liter is made, then the equivalent to 13.0 gm./l. of disodium EDTA is used.
- disodium EDTA means the disodium salt of ethylenediamine tetraacetic acid with two molecules of water of hydration, i.e., disodium EDTA dihydrate.
- CEA has a tendency to adhere to glass, it has been found necessary to use a small amount of proteinaceous material to coat the glass receptacles containing the diluent medium and prevent CEA from adhering. Such proteinaceous material must be inert with respect to the antibody-antigen reactions present when the curves are developed.
- a typical suitable material is bovine serum albumin (BSA).
- BSA bovine serum albumin
- the BSA is commercially available as a 30 percent aqueous solution and is suitable in this form for use in the invention, also BSA in the form of a dry powder is suitable.
- the amount used in forming the compositions of this invention is u liters/liter or 21 mg./ liter when dry powder is used. In the event an increase in concentration of the buffer is desired then 700 uL/l. or 210 mg./liter is used.
- the receptables are a material other than glass or glass coated with a non-reactive coating, e.g., Teflon, then there is no need for the proteinaceous material.
- a non-reactive coating e.g., Teflon
- a small amount of a preservative should be used to prevent growth or microorganisms, particularly fungi. It has been found that either 0.17 gm./l. of sodium azide or a functionally equivalent amount of other preservatives, e.g., potassium sorbate, is suitable for the compositions of this invention. In the event the more concentrated plasma extract substitute is used, then 1.7 gm./l. of preservative is required.
- the invention is not limited to the use of the specific preservatives named herein since other functionally equivalent materials or 3 mixtures thereof are suitable and would be obvious to the skilled artisan.
- the base used should be a base of the same metal forming the salt with the acid.
- 1M sodium hydroxide is used when disodium EDTA is the organic acid salt.
- the pH is adjusted similarly to about 6.25 since upon dilution it then approaches the preferred 6.5.
- the amount of base used varies depending upon the pH of the composition prior to its addition. Generally, however, approximately ml. of l M base are added.
- the composition is prepared by mixing all the ingredients except the base in about 800 ml. deionized or distilled water. Following this, the solution is brought to the preferred pH, 6.5 with the base and the solution is made to 1,000 ml. with deionized or distilled water. (In the case of a tenfold concentration increase, the pH is brought to 6.25.) The resulting solution is stable at room temperature (25C.) or 4C. for several weeks.
- carcinoembryonic antigen includes all antigenic material which is specific to antibodies of carcinoembryonic antigen.
- Typical CEA antigen materials are disclosed in US. Pat. No. 3,663,684 and US. Pat. Application Ser. No. 133,404, filed Apr. 2, 1971 now US. Pat. No. 3,697,638.
- EXAMPLE 1 Preparation of Plasma Substitute 1.3 Grams of EDTA-Na -2H O (3.7 mM) is dissolved in about 800 ml. of water. 0.17 Grams of sodium azide and 70 ul. of 30 percent aqueous BSA are added to the solution. The pH is adjusted to 6.5 with 1M sodium hydroxide and the volume made up to 1,000 with deionized or distilled water. This solution is stable at 4C. or at room temperature for several weeks.
- EXAMPLE 2 Preparation of Concentrated Plasma Substitute 13.0 Grams of EDTA-Na -2H O (37 mM) is dissolved in about 800 m1. of water. 1.7 Grams of sodium azide and 700 pl. of 30 percent aqueous BSA are added to the solution. The pH is adjusted to 6.25 with 1M sodium hydroxide and the volume made up to 1,000 ml. with deionized or distilled water. The pH of this stock solution, upon diluting tenfold to make the working solution, approaches 6.5. This stock solution is stable for several weeks at 4C. or at room temperature.
- EXAMPLE 3 Preparation of Plasma Extract 1 MI. of normal blood group Type A+ plasma is diluted with 4 ml. of saline solution in a test tube. 5 M1. of 1.2 M perchloric acid is added to each tube. The mixtures are centrifuged for 20 minutes at 1,000 X g. Thesupemates are decanted in dialysis bags which are then sealed. The bags are placed in a dialysis bath and dialyzed for 18 hours against 60 volumes of deionized or distilled water which is changed several times during the dialysis. A final 3 hour dialysis against 60 volumes of 0.01 M ammonium acetate buffer, pH 6.8 is performed. The extracts are then transferred to disposable test tubes.
- EXAMPLE 4 Antiserum Titration Curve with Plasma Extract Graded amounts, i.e., 50, 100, 200 and 500 pl of antiserum to CEA which is diluted 1 to 2,500 in a buffer comprising 9 volumes of borate buffer (pH 8.4) and 1 volume of blood group Type A+ plasma are added to four test tubes, each containing 10 ml. of plasma extract prepared as in Example 3. Only water was added to one of the test tubes for a zero measurement. The mixtures were incubated for 0.5 hour at 45C. about 3 nanograms of I CEA having 150,000-200,000 cpm. were then added to each tube and the mixture incubated for 0.5 hour at 45C. 5 M1.
- the assay indicates the activity of the antiserum, knowledge of which is needed for use in the radioimmunoassay for CEA.
- Example 5 Antiserum Titration Curve with Plasma Extract Substitute The identical procedure of Example 4 was followed except that 10 ml. of the composition of Example 1 was used in place of the 10 ml. of plasma extract. The results are shown in FIG. 1.
- EXAMPLE 6 Antiserum Titration Curve with Plasma Extract Substitute 5 ml. of the product of Example 2 were diluted to 50 ml. with deionized or distilled water and utilized as the diluent following the identical procedure of Example 5. The resulting curve is identical to that of Example 5 and shown in FIG. 1.
- EXAMPLE 7 Standard Inhibition Curve with Plasma Extract pl. of antiserum of CEA was added to each of five test tubes containing 10 ml. of the plasma extract of Example 3.
- Graded amounts of CEA i.e., 0, 2.5, 6.25, 12.5 and 25 mg, were added to the tubes and the mixture incubated for 0.5 hour at 45C.
- about 3 ng. of I -CEA with 150,000-200,000 cpm were added to each tube and the mixture incubated for 0.5 hour at 45C. 5 M].
- zirconyl phosphate gel (pH 6.25) was then added to each tube. The tubes were centrifuged at 1,000 X g for 5 minutes at room temperature.
- Example 8 Standard Inhibition Curve with Plasma Extract Substitute
- the identical procedure of Example 7 was followed except that 10 ml. of the composition of Example 1 was EXAMPLE 9 Standard Inhibition'Curve with Plasma Extract Substitute From Example 2 5 ml. of the product of Example 2 were diluted to 50 ml. with deionized or distilled water and utilized as diluent following the identical procedure of Example 8. The resulting curve is identical to that of Example 8 and shown in FIG. 2.
- the slightly higher level of radioactivity of the curve using the plasma extract at the zero value indicates the presence of a very small but measurable amount of CEA in the plasma extract.
- composition of claim 2 wherein the preservative is sodium azide.
- a diluent composition suitable for forming an antibody to CEA titration curve or a CEA standard inhibition curve substantially identical to that resulting when blood plasma is used as the diluent said composition having a pH 6.25 and comprising per liter; 13.0 grams of. disodium EDTA dihydrate, 1.7 grams of an antimi crobial preservative, 700 pl. of 30 percent w/w aqueous BSA, sodium hydroxide, and sufficient water to make 1 liter.
- composition of claim 4 wherein the preservative is sodium azide.
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Priority Applications (16)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ZA723264A ZA723264B (en) | 1972-10-13 | 1972-05-15 | Improvements in toys |
US00297565A US3852415A (en) | 1972-10-13 | 1972-10-13 | Compositions for use in radioimmunoassay as a substitute for blood plasma extract in determination of carcinoembryonic antigen |
ZA737693A ZA737693B (en) | 1972-10-13 | 1973-10-01 | Dilution agent |
CH1403573A CH584409A5 (cs) | 1972-10-13 | 1973-10-01 | |
CA182,470A CA1012458A (en) | 1972-10-13 | 1973-10-02 | Physiological buffer system |
DE2349738A DE2349738C3 (de) | 1972-10-13 | 1973-10-03 | Wäßriger Plasmaersatz |
IL43378A IL43378A (en) | 1972-10-13 | 1973-10-04 | Aqueous buffer solution for use as a plasma substitute |
IT29875/73A IT1001584B (it) | 1972-10-13 | 1973-10-08 | Agente per diluizione |
AU61242/73A AU467511B2 (en) | 1972-10-13 | 1973-10-10 | Dilution agent |
FR7336328A FR2203525A5 (cs) | 1972-10-13 | 1973-10-11 | |
DD174034A DD108151A5 (cs) | 1972-10-13 | 1973-10-12 | |
SE7313919A SE417460B (sv) | 1972-10-13 | 1973-10-12 | Sett att vid bestemning av karcinoembruonalt antigen utnyttja ett nytt spedningsmedel |
GB4780473A GB1423244A (en) | 1972-10-13 | 1973-10-12 | Dilution agent |
NL7314093.A NL161884C (nl) | 1972-10-13 | 1973-10-12 | Werkwijze voor het bepalen van carcino-embryonisch antigeen (cea) in een monster. |
BE136609A BE805991A (fr) | 1972-10-13 | 1973-10-12 | Diluant |
JP11463773A JPS5315130B2 (cs) | 1972-10-13 | 1973-10-12 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US00297565A US3852415A (en) | 1972-10-13 | 1972-10-13 | Compositions for use in radioimmunoassay as a substitute for blood plasma extract in determination of carcinoembryonic antigen |
Publications (1)
Publication Number | Publication Date |
---|---|
US3852415A true US3852415A (en) | 1974-12-03 |
Family
ID=23146844
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US00297565A Expired - Lifetime US3852415A (en) | 1972-10-13 | 1972-10-13 | Compositions for use in radioimmunoassay as a substitute for blood plasma extract in determination of carcinoembryonic antigen |
Country Status (15)
Country | Link |
---|---|
US (1) | US3852415A (cs) |
JP (1) | JPS5315130B2 (cs) |
AU (1) | AU467511B2 (cs) |
BE (1) | BE805991A (cs) |
CA (1) | CA1012458A (cs) |
CH (1) | CH584409A5 (cs) |
DD (1) | DD108151A5 (cs) |
DE (1) | DE2349738C3 (cs) |
FR (1) | FR2203525A5 (cs) |
GB (1) | GB1423244A (cs) |
IL (1) | IL43378A (cs) |
IT (1) | IT1001584B (cs) |
NL (1) | NL161884C (cs) |
SE (1) | SE417460B (cs) |
ZA (2) | ZA723264B (cs) |
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3901870A (en) * | 1974-03-12 | 1975-08-26 | Behringwerke Ag | Derivative of alpha' 1'-fetospecific serum protein and process for its manufacture |
USB554039I5 (cs) * | 1975-02-28 | 1976-02-24 | ||
US4043757A (en) * | 1975-05-20 | 1977-08-23 | Ortho Pharmaceutical Corporation | Method for detection of human mammary carcinoma |
US4132769A (en) * | 1974-10-30 | 1979-01-02 | Osther Kurt B | Cancer antigen, cancer therapy, and cancer diagnosis |
US4152410A (en) * | 1975-09-03 | 1979-05-01 | Eisai Co., Ltd. | Diagnosis reagent for neoplasm and method for diagnosis of neoplasm |
WO1981001469A1 (en) * | 1979-11-21 | 1981-05-28 | Wistar Inst | Monoclonal hybridoma antibody specific for high molecular weight carcinoembryonic antigen |
US4379839A (en) * | 1977-05-23 | 1983-04-12 | The Trustees Of Columbia University In The City Of New York | Method for detecting cancer |
US4624916A (en) * | 1984-04-06 | 1986-11-25 | International Immunoassay Laboratories, Inc. | Process and composition for the rapid quantitation of small levels of creative kinase-MB isoenzyme |
US4631254A (en) * | 1982-12-06 | 1986-12-23 | Hoffmann-La Roche Inc. | Carcinoembryonic antigen determination |
US5296377A (en) * | 1992-12-15 | 1994-03-22 | Boehringer Mannheim Corporation | Control reagent containing a hydroxylamine or an antioxidant |
US6013772A (en) * | 1986-08-13 | 2000-01-11 | Bayer Corporation | Antibody preparations specifically binding to unique determinants of CEA antigens or fragments thereof and use of the antibody preparations in immunoassays |
US10563153B2 (en) | 2010-05-20 | 2020-02-18 | Ecolab Usa Inc. | Rheology modified low foaming liquid antimicrobial compositions and methods of use thereof |
US11474114B2 (en) * | 2020-06-19 | 2022-10-18 | Republic of Korea (National Forensic Service Director Ministry of the Interior and Safety) | Artificial blood for bloodstain pattern analysis |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS562557A (en) * | 1979-06-22 | 1981-01-12 | Green Cross Corp:The | Aqueous solvent for coagulation test |
JPS57208459A (en) * | 1981-06-19 | 1982-12-21 | Eisai Co Ltd | Measuring method using enzyme-labelled antibody and reagent |
JPS61280566A (ja) * | 1984-09-03 | 1986-12-11 | Hiroaki Sawai | 2’−5’−オリゴアデニル酸の免疫学的直接定量法 |
US4690772A (en) * | 1985-06-03 | 1987-09-01 | National Medical Care | Sterilant compositions |
CA1296253C (en) * | 1986-10-20 | 1992-02-25 | Praveen Tyle | Stabilized growth hormone compositions |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3697638A (en) * | 1970-06-01 | 1972-10-10 | Hoffmann La Roche | Antigens |
US3720760A (en) * | 1968-09-06 | 1973-03-13 | Pharmacia Ab | Method for determining the presence of reagin-immunoglobulins(reagin-ig)directed against certain allergens,in aqueous samples |
-
1972
- 1972-05-15 ZA ZA723264A patent/ZA723264B/xx unknown
- 1972-10-13 US US00297565A patent/US3852415A/en not_active Expired - Lifetime
-
1973
- 1973-10-01 ZA ZA737693A patent/ZA737693B/xx unknown
- 1973-10-01 CH CH1403573A patent/CH584409A5/xx not_active IP Right Cessation
- 1973-10-02 CA CA182,470A patent/CA1012458A/en not_active Expired
- 1973-10-03 DE DE2349738A patent/DE2349738C3/de not_active Expired
- 1973-10-04 IL IL43378A patent/IL43378A/en unknown
- 1973-10-08 IT IT29875/73A patent/IT1001584B/it active
- 1973-10-10 AU AU61242/73A patent/AU467511B2/en not_active Expired
- 1973-10-11 FR FR7336328A patent/FR2203525A5/fr not_active Expired
- 1973-10-12 BE BE136609A patent/BE805991A/xx not_active IP Right Cessation
- 1973-10-12 NL NL7314093.A patent/NL161884C/xx not_active IP Right Cessation
- 1973-10-12 DD DD174034A patent/DD108151A5/xx unknown
- 1973-10-12 SE SE7313919A patent/SE417460B/xx unknown
- 1973-10-12 GB GB4780473A patent/GB1423244A/en not_active Expired
- 1973-10-12 JP JP11463773A patent/JPS5315130B2/ja not_active Expired
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3720760A (en) * | 1968-09-06 | 1973-03-13 | Pharmacia Ab | Method for determining the presence of reagin-immunoglobulins(reagin-ig)directed against certain allergens,in aqueous samples |
US3720760B1 (cs) * | 1968-09-06 | 1984-02-07 | Pharmacia Ab | |
US3697638A (en) * | 1970-06-01 | 1972-10-10 | Hoffmann La Roche | Antigens |
Non-Patent Citations (3)
Title |
---|
Aach et al., Proc. Nat. Acad. Sci., U.S.A., Vol. 68, No. 5, p. 1056, May 1971. * |
Salmon et al., The Journal of Immunology, Vol. 103, No. 1, July 1969, pp. 129 131. * |
Salmon et al., The Journal of Immunology, Vol. 104, No. 3, March 1970, pp. 665 667. * |
Cited By (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3901870A (en) * | 1974-03-12 | 1975-08-26 | Behringwerke Ag | Derivative of alpha' 1'-fetospecific serum protein and process for its manufacture |
US4132769A (en) * | 1974-10-30 | 1979-01-02 | Osther Kurt B | Cancer antigen, cancer therapy, and cancer diagnosis |
USB554039I5 (cs) * | 1975-02-28 | 1976-02-24 | ||
US3999944A (en) * | 1975-02-28 | 1976-12-28 | Hoffmann-La Roche Inc. | Detection of breast cancer |
US4043757A (en) * | 1975-05-20 | 1977-08-23 | Ortho Pharmaceutical Corporation | Method for detection of human mammary carcinoma |
US4152410A (en) * | 1975-09-03 | 1979-05-01 | Eisai Co., Ltd. | Diagnosis reagent for neoplasm and method for diagnosis of neoplasm |
US4379839A (en) * | 1977-05-23 | 1983-04-12 | The Trustees Of Columbia University In The City Of New York | Method for detecting cancer |
US4349528A (en) * | 1979-11-21 | 1982-09-14 | The Wistar Institute | Monocolonal hybridoma antibody specific for high molecular weight carcinoembryonic antigen |
WO1981001469A1 (en) * | 1979-11-21 | 1981-05-28 | Wistar Inst | Monoclonal hybridoma antibody specific for high molecular weight carcinoembryonic antigen |
US4631254A (en) * | 1982-12-06 | 1986-12-23 | Hoffmann-La Roche Inc. | Carcinoembryonic antigen determination |
US4624916A (en) * | 1984-04-06 | 1986-11-25 | International Immunoassay Laboratories, Inc. | Process and composition for the rapid quantitation of small levels of creative kinase-MB isoenzyme |
US6013772A (en) * | 1986-08-13 | 2000-01-11 | Bayer Corporation | Antibody preparations specifically binding to unique determinants of CEA antigens or fragments thereof and use of the antibody preparations in immunoassays |
US6022958A (en) * | 1986-08-13 | 2000-02-08 | Bayer Corporation | cDNAs coding for members of the carcinoembryonic antigen family |
US5296377A (en) * | 1992-12-15 | 1994-03-22 | Boehringer Mannheim Corporation | Control reagent containing a hydroxylamine or an antioxidant |
US10563153B2 (en) | 2010-05-20 | 2020-02-18 | Ecolab Usa Inc. | Rheology modified low foaming liquid antimicrobial compositions and methods of use thereof |
US11268049B2 (en) | 2010-05-20 | 2022-03-08 | Ecolab Usa Inc. | Rheology modified low foaming liquid antimicrobial compositions and methods of use thereof |
US12252672B2 (en) | 2010-05-20 | 2025-03-18 | Ecolab Usa Inc. | Rheology modified low foaming liquid antimicrobial compositions and methods of use thereof |
US11474114B2 (en) * | 2020-06-19 | 2022-10-18 | Republic of Korea (National Forensic Service Director Ministry of the Interior and Safety) | Artificial blood for bloodstain pattern analysis |
Also Published As
Publication number | Publication date |
---|---|
IL43378A0 (en) | 1974-01-14 |
DE2349738A1 (de) | 1974-04-18 |
GB1423244A (en) | 1976-02-04 |
DD108151A5 (cs) | 1974-09-05 |
DE2349738B2 (de) | 1978-01-19 |
AU467511B2 (en) | 1975-12-04 |
BE805991A (fr) | 1974-04-12 |
JPS5315130B2 (cs) | 1978-05-23 |
NL7314093A (cs) | 1974-04-16 |
NL161884C (nl) | 1980-03-17 |
IL43378A (en) | 1976-04-30 |
FR2203525A5 (cs) | 1974-05-10 |
IT1001584B (it) | 1976-04-30 |
JPS4971133A (cs) | 1974-07-10 |
ZA723264B (en) | 1974-08-28 |
DE2349738C3 (de) | 1981-06-19 |
AU6124273A (en) | 1975-05-01 |
SE417460B (sv) | 1981-03-16 |
ZA737693B (en) | 1974-08-28 |
NL161884B (nl) | 1979-10-15 |
CH584409A5 (cs) | 1977-01-31 |
CA1012458A (en) | 1977-06-21 |
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