US3548051A - Preparation of erythrocytes for immunochemical determination of human chorionic gonadotropin - Google Patents
Preparation of erythrocytes for immunochemical determination of human chorionic gonadotropin Download PDFInfo
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- US3548051A US3548051A US623825A US3548051DA US3548051A US 3548051 A US3548051 A US 3548051A US 623825 A US623825 A US 623825A US 3548051D A US3548051D A US 3548051DA US 3548051 A US3548051 A US 3548051A
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- United States
- Prior art keywords
- erythrocytes
- hcg
- preparation
- human chorionic
- immunochemical
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Links
- 210000003743 erythrocyte Anatomy 0.000 title description 52
- 102000011022 Chorionic Gonadotropin Human genes 0.000 title description 29
- 108010062540 Chorionic Gonadotropin Proteins 0.000 title description 29
- 229940084986 human chorionic gonadotropin Drugs 0.000 title description 29
- 230000000984 immunochemical effect Effects 0.000 title description 13
- 238000002360 preparation method Methods 0.000 title description 7
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 44
- 238000000034 method Methods 0.000 description 14
- 239000000725 suspension Substances 0.000 description 13
- 239000000427 antigen Substances 0.000 description 12
- 102000036639 antigens Human genes 0.000 description 12
- 108091007433 antigens Proteins 0.000 description 12
- 206010070834 Sensitisation Diseases 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000843 powder Substances 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000004615 ingredient Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 235000018553 tannin Nutrition 0.000 description 4
- 229920001864 tannin Polymers 0.000 description 4
- 239000001648 tannin Substances 0.000 description 4
- 210000002700 urine Anatomy 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 3
- 230000004520 agglutination Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- -1 for instance Proteins 0.000 description 3
- 239000012266 salt solution Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 102000006771 Gonadotropins Human genes 0.000 description 2
- 108010086677 Gonadotropins Proteins 0.000 description 2
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 239000000783 alginic acid Substances 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229960001126 alginic acid Drugs 0.000 description 2
- 150000004781 alginic acids Chemical class 0.000 description 2
- 235000013681 dietary sucrose Nutrition 0.000 description 2
- LEQAOMBKQFMDFZ-UHFFFAOYSA-N glyoxal Chemical compound O=CC=O LEQAOMBKQFMDFZ-UHFFFAOYSA-N 0.000 description 2
- 239000002622 gonadotropin Substances 0.000 description 2
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 230000035935 pregnancy Effects 0.000 description 2
- 238000009597 pregnancy test Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 229960004793 sucrose Drugs 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 241000283086 Equidae Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 102000009151 Luteinizing Hormone Human genes 0.000 description 1
- 108010073521 Luteinizing Hormone Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-N PYRUVIC-ACID Natural products CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- XUGUHTGSMPZQIW-UHFFFAOYSA-N [[4-(4-diazonioiminocyclohexa-2,5-dien-1-ylidene)cyclohexa-2,5-dien-1-ylidene]hydrazinylidene]azanide Chemical compound C1=CC(N=[N+]=[N-])=CC=C1C1=CC=C(N=[N+]=[N-])C=C1 XUGUHTGSMPZQIW-UHFFFAOYSA-N 0.000 description 1
- 230000000274 adsorptive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000007900 aqueous suspension Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 229960004407 chorionic gonadotrophin Drugs 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 229960001484 edetic acid Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 108010074605 gamma-Globulins Proteins 0.000 description 1
- 229940015043 glyoxal Drugs 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000008240 homogeneous mixture Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229940040129 luteinizing hormone Drugs 0.000 description 1
- 229940057948 magnesium stearate Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 229960001855 mannitol Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000021317 phosphate Nutrition 0.000 description 1
- 150000003013 phosphoric acid derivatives Chemical class 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 229920001592 potato starch Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000012475 sodium chloride buffer Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- IBUIVNCCBFLEJL-UHFFFAOYSA-M sodium;phosphoric acid;chloride Chemical compound [Na+].[Cl-].OP(O)(O)=O IBUIVNCCBFLEJL-UHFFFAOYSA-M 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229960000814 tetanus toxoid Drugs 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/554—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being a biological cell or cell fragment, e.g. bacteria, yeast cells
- G01N33/555—Red blood cell
- G01N33/556—Fixed or stabilised red blood cell
-
- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
- G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors
-
- 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
- Y10S436/815—Test for named compound or class of compounds
- Y10S436/817—Steroids or hormones
- Y10S436/818—Human chorionic gonadotropin
Definitions
- erythrocytes from mammals such as cattle, horses, sheep, rabbits, as well as chickens and from human beings, can be used as an auxiliary in immunochemical determinations based on the reaction of an antigen with its antibody.
- the erythrocytes are applied, for instance, as carrier of antigens, for instance, human chorionic gonadotropin (HCG), serum gonadotropin, growth hormone, and further serum albumin, gamma globulins, insulin, luteinizing hormone and tetanus toxoid.
- HCG human chorionic gonadotropin
- serum gonadotropin serum gonadotropin
- growth hormone growth hormone
- serum albumin serum albumin
- gamma globulins insulin, luteinizing hormone and tetanus toxoid.
- tanning agents or mordants to improve the adsorptive properties of the erythrocytes is described in the literature: inulin, tannin, hydroquinone, bis-diazobenzidine, Formalin, acetaldehyde, pyruvic acid aldehyde and glyoxal. Also successive treatment of the erythrocytes with different tanning agents and mordants has already been applied, for example, a pre-treatment with Formalin, followed by reaction with tannin.
- erythrocytes sensitised after the pre-treatment have been stored as an aqueous suspension, but preferably in a freeze-dried form.
- erythrocytes are very unstable particles and the preservation of these carriers has already required much study.
- the erythrocytes pretreated with a mordant are less unstable and can be readily freeze-dried, while in a freeze-dried state they are stable for a pretty long time, There exists a need of a presentation form, however, in which the erythrocytes can be well handled and are at the same time stable and have kept all their properties essential for the performance of immunochemical determination methods.
- HCG human chorionic gonadotrophin
- Erythrocytes treated according to the invention have become strong enough to be compressed to, say, tablets.
- the conventional auxiliaries can be applied, which must not be aggressive against erythrocytes and the antigen attached to them. It stands to reason that the bond between the antigen and the erythrocytes must not be affected adversely by the auxiliaries used and that they must not have a perceptible influence on the determinations for which the erythrocytes are intended, so that the sensibility, the accuracy and the velocity of the determination reaction is not essentially changed.
- erythrocytes treated according to the invention do not only have greatly improved mechanical properties, but also a greater sensibility to agglutination with antiserum.
- complete agglutination could even have been obtained with a nontreated suspension of erythrocytes sensitised with HCG with a dilution of antiserum of while after treatment in accordance with the invention the same suspension caused the same reaction with a dilution of antiserum of from /7500 to A5000-
- the invention is not limited to application in the sensitisation of erythrocytes with HCG; it can also be applied in the sensitisation with any antigen that can stand up to the present treatment with formaldehyde.
- both the powder of sensitised erythrocytes obtained, for example, after freeze-drying or spray'drying of the suspension, can be applied and the tablets compressed therefrom.
- the powders obtained after freeze-drying of suspensions of erythrocytes are very hygroscopic, they are mixed and compressed with the other auxiliaries, preferably in an atmosphere with a slight relative degree of moisture, preferably to about It is advantageous to choose such ingredients and apply such conditions in compressing that the disintegration time of the tablets is short, for example, less than 3 minutes.
- auxiliaries to be applied are amongst others usable; saccharose, manitol, lactose and ureum; as lubricants for example: boric acid, starch, Carboway 4000 (polyethylene glycol) and magnesium stearate, as disintegration agents for example: starch and alginic acid.
- ingredients are for instance substances used in the said determination, such as phosphates, for example, monopotassium phosphate and disodium phosphate, serving as buffers for the suspension prepared from the tablets, further sodium chloride and a chelating agent, such as the disodium salt of ethylene-diamine tetra acetic acid (E.D.T.A.) and albumin.
- phosphates for example, monopotassium phosphate and disodium phosphate
- a chelating agent such as the disodium salt of ethylene-diamine tetra acetic acid (E.D.T.A.) and albumin.
- the required ingredients can be mixed in a dry condition with the treated erythrocytes before tabletting, but it is more advantageous to dissolve the soluble ingredients together and to dry them by spray-drying, followed by mixing with the treated erythrocytes to obtain a very homogeneous mixture and to prevent the erythrocytes from being damaged during the further treatment by coarse crystalline components of the auxiliaries.
- insoluble auxiliaries In the processing of insoluble auxiliaries special attention should be paid that they do not disturb the determinations performed with a suspension obtained from the tablets. For the insoluble components will be able easily to change the sedimentation pattern of the erythrocytes, thus influencing the accuracy of the observation unfavourably.
- EXAMPLE 1 A suspension of sheep erythrocytes treated in a known manner with Formalin and tannin was centrifuged and washed with a phosphate sodium chloride buffer of pH 6.4 and next incorporated in this buffer. To this mixture an equal volume was added of a solution of 50 I.U. HCG per ml. in a buffer of pH 6.4. This mixture was stored for 48 hours at 37 C., whereupon formaldehyde was added till a final concentration had been obtained of 0.25% (weight/volume) formaldehyde. Next this mixture was stored for 15 hours at a temperature of 37 C., whereupon the erythrocytes were again centrifuged, washed with a physiological salt solution, incorporated as 10% v./v. suspension in a physiological salt solution containing 0.1% of bovine serum albumin, and stored for a few days at 04 C.
- the powder is a good reagent on HCG.
- EXAMPLE II 300 ml. of the suspension of erythrocytes of pH 6.4 according to Example I were treated with 300 ml. of a solution of HCG with 50 I.U. HCG per ml. at pH 7.6 and next with formaldehyde in a final concentration of 0.5%. After freeze-drying the powder was tabletted. The tablets obtained gave excellent results as a reagent in immunochemical pregnancy tests.
- EXAMPLE III By the method of Example I erythrocytes were sensitised with HCG in the presence of formaldehyde in a final concentration of 0.25%, and that for the last 24 hours at 30 C. After freeze-drying and compression tablets were obtained which gave excellent results as a reagent in an HCG test.
- EXAMPLE IV Experiments were performed with treatment with formaldehyde for the last 15 hours of the sensitisation of erythrocytes with HCG in accordance with Example I, but this time at pH 5.5.
- the suspension was dried in a Niro spray-drier with an inlet temperature of C. and an outlet temperature of 75 C.
- the resulting powder was next mixed with 0.4% (weight-weight) potato starch and compressed to tablets of about 15 mg. They gave excellent results as a reagent in immunochemical pregnancy determinations.
- sensitized erythrocytes are admixed with an auxiliary for tableting selected from the group consisting of saccharose, mannitol, magnesium stearate, and alginic acid, and then compressed into tablet form.
- an auxiliary for tableting selected from the group consisting of saccharose, mannitol, magnesium stearate, and alginic acid
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Endocrinology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Reproductive Health (AREA)
- Mycology (AREA)
- Medicinal Preparation (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| NL6603909A NL6603909A (enrdf_load_stackoverflow) | 1966-03-24 | 1966-03-24 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3548051A true US3548051A (en) | 1970-12-15 |
Family
ID=19796083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US623825A Expired - Lifetime US3548051A (en) | 1966-03-24 | 1967-03-17 | Preparation of erythrocytes for immunochemical determination of human chorionic gonadotropin |
Country Status (12)
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3862302A (en) * | 1969-03-20 | 1975-01-21 | Akzona Inc | Pelletized pregnancy test reagents |
| US3987159A (en) * | 1973-03-02 | 1976-10-19 | Schering Aktiengesellschaft | Stable sensitized erythrocytes and preparation means |
| US4136161A (en) * | 1976-03-16 | 1979-01-23 | Ortho Diagnostics, Inc. | Stabilized erythrocytes and methods therefor |
| US4282002A (en) * | 1979-09-06 | 1981-08-04 | Akzona Incorporated | Sensitized sheep stroma immunoassay for rheumatoid factor |
| FR2543300A1 (fr) * | 1983-03-23 | 1984-09-28 | Yanovsky Jorge | Reactif serologique |
| US4587222A (en) * | 1980-02-15 | 1986-05-06 | Laboratories Polypharma | Reagent comprising treated red blood cells and methods for detecting rheumatoid factor |
| US4690908A (en) * | 1979-06-05 | 1987-09-01 | Mochida Seiyaku Kabushiki Kaisha | Measuring reagent kit for determining an antigen, antibody or antigen-antibody complex |
| US20070178434A1 (en) * | 2004-02-02 | 2007-08-02 | I.M.T. Interface Multigrad Technology Ltd. | Biological material and methods and solutions for preservation thereof |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5045446A (en) * | 1988-08-26 | 1991-09-03 | Cryopharm Corporation | Lyophilization of cells |
| IL90188A0 (en) * | 1988-05-18 | 1989-12-15 | Cryopharm Corp | Process and medium for the lyophilization of erythrocytes |
| EP0356258A3 (en) * | 1988-08-26 | 1990-06-06 | Cryopharm Corporation | Processes for the lyophilization of red blood cells, together with media for lyophilization |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3096250A (en) * | 1959-11-30 | 1963-07-02 | Indiana University Foundation | Novel particulate antigens and process |
| US3236732A (en) * | 1962-01-22 | 1966-02-22 | Edward R Arquilla | Pregnancy test method and immunological indicator therefor |
-
0
- NL NL126572D patent/NL126572C/xx active
-
1966
- 1966-03-24 NL NL6603909A patent/NL6603909A/xx unknown
-
1967
- 1967-03-12 IL IL27578A patent/IL27578A/xx unknown
- 1967-03-17 US US623825A patent/US3548051A/en not_active Expired - Lifetime
- 1967-03-18 ES ES338201A patent/ES338201A1/es not_active Expired
- 1967-03-20 DE DE19671617672 patent/DE1617672A1/de active Pending
- 1967-03-21 CH CH412467A patent/CH508390A/de not_active IP Right Cessation
- 1967-03-22 DK DK155467AA patent/DK126452B/da unknown
- 1967-03-22 GB GB03515/67A patent/GB1179131A/en not_active Expired
- 1967-03-22 GR GR670136578A patent/GR36578B/el unknown
- 1967-03-23 AT AT284467A patent/AT270864B/de active
- 1967-03-23 SE SE04108/67A patent/SE335592B/xx unknown
- 1967-03-24 BE BE696050D patent/BE696050A/xx unknown
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3096250A (en) * | 1959-11-30 | 1963-07-02 | Indiana University Foundation | Novel particulate antigens and process |
| US3236732A (en) * | 1962-01-22 | 1966-02-22 | Edward R Arquilla | Pregnancy test method and immunological indicator therefor |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3862302A (en) * | 1969-03-20 | 1975-01-21 | Akzona Inc | Pelletized pregnancy test reagents |
| US3987159A (en) * | 1973-03-02 | 1976-10-19 | Schering Aktiengesellschaft | Stable sensitized erythrocytes and preparation means |
| US4136161A (en) * | 1976-03-16 | 1979-01-23 | Ortho Diagnostics, Inc. | Stabilized erythrocytes and methods therefor |
| US4690908A (en) * | 1979-06-05 | 1987-09-01 | Mochida Seiyaku Kabushiki Kaisha | Measuring reagent kit for determining an antigen, antibody or antigen-antibody complex |
| US4282002A (en) * | 1979-09-06 | 1981-08-04 | Akzona Incorporated | Sensitized sheep stroma immunoassay for rheumatoid factor |
| US4587222A (en) * | 1980-02-15 | 1986-05-06 | Laboratories Polypharma | Reagent comprising treated red blood cells and methods for detecting rheumatoid factor |
| FR2543300A1 (fr) * | 1983-03-23 | 1984-09-28 | Yanovsky Jorge | Reactif serologique |
| US7935478B2 (en) * | 2004-02-02 | 2011-05-03 | Core Dynamics Limited | Biological material and methods and solutions for preservation thereof |
| US20070178434A1 (en) * | 2004-02-02 | 2007-08-02 | I.M.T. Interface Multigrad Technology Ltd. | Biological material and methods and solutions for preservation thereof |
| US20110177488A1 (en) * | 2004-02-02 | 2011-07-21 | Core Dynamics Limited | Biological material and methods and solutions for preservation thereof |
| US8512941B2 (en) | 2004-02-02 | 2013-08-20 | Core Dynamics Limited | Biological material and methods and solutions for preservation thereof |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11609042B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11609043B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11740019B2 (en) | 2019-03-14 | 2023-08-29 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11747082B2 (en) | 2019-03-14 | 2023-09-05 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11994343B2 (en) | 2019-03-14 | 2024-05-28 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
Also Published As
| Publication number | Publication date |
|---|---|
| CH508390A (de) | 1971-06-15 |
| AT270864B (de) | 1969-05-12 |
| NL6603909A (enrdf_load_stackoverflow) | 1967-09-25 |
| IL27578A (en) | 1971-04-28 |
| GB1179131A (en) | 1970-01-28 |
| DE1617672A1 (de) | 1971-04-01 |
| GR36578B (el) | 1969-02-21 |
| DK126452B (da) | 1973-07-16 |
| ES338201A1 (es) | 1968-04-01 |
| NL126572C (enrdf_load_stackoverflow) | |
| BE696050A (enrdf_load_stackoverflow) | 1967-09-25 |
| SE335592B (enrdf_load_stackoverflow) | 1971-06-01 |
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