DE2262413A1 - Direkte radioimmunpruefung auf antigene und ihre antikoerper - Google Patents

Direkte radioimmunpruefung auf antigene und ihre antikoerper

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DE2262413A1
DE2262413A1 DE2262413A DE2262413A DE2262413A1 DE 2262413 A1 DE2262413 A1 DE 2262413A1 DE 2262413 A DE2262413 A DE 2262413A DE 2262413 A DE2262413 A DE 2262413A DE 2262413 A1 DE2262413 A1 DE 2262413A1
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antigen
coating
antibody
solution
buffer mixture
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DE2262413B2 (de
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Chung-Mei Ling
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Abbott Laboratories
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    • 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
    • G01N33/54306—Solid-phase reaction mechanisms
    • 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
    • G01N33/576—Immunoassay; Biospecific binding assay; Materials therefor for hepatitis
    • 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
    • Y10S436/804—Radioisotope, e.g. radioimmunoassay
    • 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
    • Y10S436/82—Hepatitis associated antigens and antibodies

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Pathology (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Communicable Diseases (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Description

Die Erfindung betrifft ein diagnostisches Verfahren zur
Durchführung einer Radioimmunprüfung auf Antigene und ihre Antikörper, sowie ein Verfahren zur Herstellung eines Überzuges bei einer für diese Prüfung verwendbaren Vorrichtung. Insbesondere batrifft die Erfindung ein direktes Verfahren zur Bestimmung von Hepatitis vergesellschafteten Antigenen und ihren Antikörpern sowie ein Verfahren zur Herstellung einer hierfür verwendbaren diagnostischen Vorrichtung.
Es sind zwar schon eine Anzahl Verfahren zur Peststellung der Anwesenheit von antigen-aktiven Makromolekülen, wie
intakten Viren, Viruskapsiden (virus capsids), Virusuntereinheiten, Bakterien, Membranen, Zellwänden, Hormonen usw.,
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bekannt, jedoch fehlte es bisher an einer einfachen und doch empfindlichen Prüfmethode und -vorrichtung zur Feststellung der Anwesenheit dieser Materialien. Virushepatitis, einschließlich der sogenannten Serumhepatitis, die eine verhältnismäßig häufig auftretende Krankheit ist, konnte bisher nicht leicht durch einen Test, der sowohl spezifisch als auch reproduzierbar ist und eine rasche Feststellung, ob das Serum eines Patienten oder eines Spenders Hepatitis vergesellschaftete Antigene oder Antikörper enthält oder nicht, diagnostiziert werden.
Für verschiedene Antigen/Antikörper-Materialien sind schon Radioimmunprüfmethoden entwickelt worden. Diese Prüfmethoden, wie sie beispielsweise von Kevin Catt et al in "journal of Biochemistry", I966, Band 100, S. 31c und 33c und in "Science", Band I58, S, 1570, 1967, beschrieben sind, sind indirekte Prüfungen, bei denen die anwesende Menge an Antigen der von dem Tracer emittierten Strahlung etwa umgekehrt proportional ist. Diese Prüfmethoden erforderten die Verwendung von Korrelationstabellen und anderen Materialien, so daß die Ergebnisse schlecht reproduzierbar und ungenau waren.
Durch die vorliegende Erfindung v/erden diese Schwierigkeiten, d.h. der Mangel an Reproduzierbarkeit und Genauigkeit einer solchen Untersuchung, überwunden. Das Verfahren gemäß der Erfindung besteht darin, daß man eine Probe eines unbekannten Serums mit einer Vorrichtung, die mit einem Antikörper-Überzug versehen ist, in Kontakt bringt, die Prüfvorrichtung und das Serum 1/2 bis h2 Stunden bebrütet, absaugt und wäscht, einen J-125 als Tracer enthaltenden Antikörper mit dem Serum und der mit dem Überzug versehenen Vorrichtung in Kontakt bringt und 1 bis 6 Stunden bebrütet, absaugt und wäscht und die J-125-Gamma-Strahlung der Vorrichtung auszählt.
Hauptaufgabe der Erfindung ist also ein neues Verfahren zur direkten Bestimmung von Antigenen und ihren Antikörpern.
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Eine weitere Aufgabe der Erfindung ist ein Verfahren zur Herstellung eines Überzuges auf einer diagnostischen Vorrichtung für eine Verwendung bei Radioimmunbestimmungen.
Eine weitere Aufgabe der Erfindung ist ein Verfahren zur raschen und genauen Bestimmung der Anwesenheit von Hepatitis vergesellschafteten Antigenen oder Antikörpern im Serum.
Die Zeichnung zeigt eine mit einem Überzug versehene Prüfvorrichtung für die Durchführung des Verfahrens gemäß der Erfindung im Schnitt.
Die Vorrichtung 1 hat die Form eines Teströhrchens mit .einem mit einem Überzug versehenen Teil 2. Dieser Teil 2 besitzt einen Überzug aus einem Antigen oder seinem Antikörper, vorzugsweise, wie gezeigt, im Bodenteil des Röhrchens. Das Verfahren gemäß der Erfindung kann aber auch mit anders als in der Zeichnung gezeigt ausgebildeten Vorrichtungen durchgeführt ■ werden.
Wie erwähnt, weist der Teil 2 der Vorrichtung 1 einen Überzug aus entweder einem Antigen oder einem Antikörper auf, je nachdem, was für ein Material untersucht werden soll. Das Verfahren gemäß der Erfindung ist auf fast alle Antigene und Antikörper in gleicher Weise anwendbar, und die Herstellung des Überzuges soll im folgenden unter Bezugnahme auf einen bestimmten Hepatitis vergesellschafteten Antikörper, d.h. Anti-Australien-Antigen (anti-Australia antigen) , beschrieben werden. Eine Anti-Australien-Antigen -Lösung mit einer Konzentration von etwa 1 bis etwa 100 μg Eiweiß/ml wird aus einem Anti-Australien-Antigen-Serum in etwa 0,005 bis etwa 0,02m 2-Amino-2-hydröxymethyl-:l. ,^-propandiol-HCl (Tris-HCl) hergestellt. Das Tris-HCl die.nt als Puffer zur Einstellung des' pH der Lösung auf etwa 7,1 bis etwa 9,5 zusammen mit etwa 0,01$ bis etwa 0,0'j/j Natriumazid. Mit dieser Anti-Australien-Antigen-Lösung wird dann der Überzug auf die Oberfläche des Röhrchens aufgebracht und 6 bis 72 Stunden, vor-
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zugsweise 12 bis 48 Stunden, bei Raumtemperatur bebrütet. Die mit dem überzug versehenen Röhrchen werden dann mit etwa 0,(005 bis etwa 0,02m Tris-HCl vom pH 6,9 bis 8,4 plus etwa 0,01% bis etwa 0,05/6 Natriumazid gewaschen. Nach diesem Waschen und Spülen können die TestrÖhrchen bei 40C aufbewahrt werden, bis sie für eine Radiοimmunprüfung gebraucht werden.
Vorzugsweise wird sowohl für das Inkubationsmedium als auch die Waschflüssigkeit eine auf pH 7,1 eingestellte 0;01m Lösung von Tris-HCl und 0,02$ Natriumazid verwendet.
Die Menge an Antikörper oder Antigen, die in dem überzug des Röhrchens anwesend ist, ist nicht kritisch, weil der Test jedesmal im Vergleich mit wenigstens einem Leertest durchgeführt wird. Für die Auswertung des Tests gemäß der Erfindung sind keine genormten Kurven oder Auftragungen erforderlich. D.h. in dem Überzug muß keine bestimmte Menge an Antikörper oder Antigen anwesend sein, sofern zwei gleiche Röhrchen verwendet werden.
Der überzug muß natürlich nicht auf ein TestrÖhrchen, sondern kann auch beispielsweise auf einen Einsatz für einen Schacht usw. aufgebracht werden, indem man den Einsatz in die Antigen- oder Antikörper-Lösung eintaucht und im übrigen nach dem oben beschriebenen Verfahren verfährt.
Antigene und Antikörper, die nach dem Verfahren gemäß der Erfindung bestimmt werden können, sind beispielsweise: verschiedene intakte Viren, Viruskapsiden, Virusuntereinheiten, Bakterien, Membranen, ZeJlwände, verschiedene Hormone, Gammaglobuline usw. Wesentlich ist nur, daß diese Materialien mindestens zwei antigen-aktive Stellen besitzen. Außerdem sind Antigene und Antikörper mit mehreren kombinierenden Stellen auch in Anwesenheit der entsprechenden Antikörper und Antigene auffindbar, sofern wenigstens zwei freie kombinierende Stellen verfügbar bleiben. Der Radioirnmuntest gemäß der Er-
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findung eignet sich zwar zur Auffindung aller obigen Materialien, insbesondere aber zur Peststellung der Anwesenheit von Hepatitis vergesellschafteten Antigenen und Antikörpern, wie Australien-Antigen und Anti-Australien-Antigen, und dieser Test stellt daher die bevorzugte Durchführungsform des Verfahrens gemäß der Erfindung dar.
Das Verfahren gemäß der Erfindung wird im folgenden für bestimmte Materialien und die zur Durchführung des direkten Radioimmuntests zur Peststellung der Anwesenheit von Hepatitis vergesellschafteten Australien-Antigen beschrieben»
Zunächst wird eine abgemessene Probe Plasma oder Blut, die auf Hepatitis vergesellschaftetes Antigen untersucht werden soll, in ein Teströhrehen mit einem Anti-Australien-Antigeη-Überzug eingebracht» Das Material wird 1/2 bis 52 Stunden, vorzugsweise 12 bis 24 Stunden, bei Raumtemperatur bebrütet und dann mit dem Puffergemisch, d.h. Tris-HCl und Natriumazid, gewaschen. Dann wird dem Röhrchen oder dem Aufnahmeschacht eine abgemessene Menge Anti-Australien-Antigen mit gereinigtem J-I25 als Tracer zugesetzt, und das damit in Kontakt stehende Röhrchen oder der Einsatz wird für weitere 1 bis 24 Spunden, vorzugsweise 1 1/2 bis 6 Stunden, bei Raumtemperatur bebrütet. Danach wird der Inhalt mit dem Puffergemisch gewaschen, und das Röhrchen wird in einen Zähler, der Gamma-Strahlung nachzuweisen vermag, eingesetzt. Gleichzeitig werden in gleicher Weise Vergleichstests unter Verwendung eines normalen Plasma statt des zu untersuchenden durchgeführt. Wenn das unbekannte Plasma■. einen höheren Gamma-Strahlungswert ergibt als die Vergleichsprobe, wird es als hepatitis-antigen-positiv angesehen.
Im allgemeinen erfolgt die Auszählung; vorzugsweise für 1 Minute. Wenn jedoch eine Probe der oberen Grenze der Vergleiohsprobe sehr nahe kommt, kann eine längere Auszählung bis zu 10 Minuten angewandt werden, damit genaue Ergebnisse erhalten werden»
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Wie oben erwähnt, erfolgt die Bebrütung Im allgemeinen bei Raumtemperatur. Um die Bebrütungszeit abzukürzen, kann jedoch leicht, d.h. bis zu etwa 550C* erwärmt werden.
Das Puffermedium enthält im allgemeinen etwa 0,005 bis etwa 0,02m Tris-HCl und etwa 0,01 bis 0,05 Gew.-^ Natriumazid bei einem pH von 6,9 bis 8,4. Der bevorzugte Puffer enthält 0,01m Tris-HCl und 0,02 Gew.-% Natriumazid.
Im allgemeinen wird der Test mit unverdünntem Blutserum oder Plasma durchgeführt. Wenn jedoch nur begrenzt Proben zur Verfügung stehen, kann die Probe in geeigneter Verdünnung mit normalem Serum oder Plasma, wie RInderserumalbumen oder einem Puffergemisch, wie einem Gemisch von Tris-HCl und Natriumazid, einem Gemisch von Tris-HCl, Natriumazid und X% Hinderserumalbumen usw., verwendet werden.
Auch kann statt des bevorzugten radioaktiven Materials J-125 auch jedes andere allgemein als Tracer für Antigene oder Antikörper in Radioimmuntests verwendete radioaktive Isotop verwendet werden.
Wie erwähnt, ist das Verfahren gemäß der Erfindung ein direkter Radioimmuntest zur Feststellung verschiedener Antigene und ihrer Antikörper, insbesondere des Hepatitis vergesellschafteten Antigens oder seines Antikörpers. Wenn das Hepatitis vergesellschaftete Antigen quantitativ bestimmt werden soll, kann eine genormte Kurve, die direkt die Beziehung zwischen den Auszählungen je Minute und der Menge an Hepatitis vergesellschaftetem Antigen wiedergibt, verwendet werden.
Das folgende Beispiel veranschaulicht die Erfindung. Angaben in Teilen und Prozent beziehen sich auf das Gewicht.
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Beispiel
Das in der Zeichnung gezeigte Röhrchen, das aus Polystyrol besteht, wird mit einem gereinigten Hepatitis vergesellschafteten Antikörper überzogen. Dieser Überzug wird aufgebracht, indem man die Oberfläche des Röhrchens einer verdünnten Lösung von Hepa- .-titis vergesellschaftetem Antikörper in 0,01m Tris-HPl vom pH 7,1 und 0,02 Gew.-% Natriumazid aussetzt, und das'mit dem Überzug versehene Röhrchen «wird dann 1 Tag bei Raumtemperatur bebrütet. Dann wird das Röhrchen mit (aliquots of) 0,Ölm Tris-HCl plus 0,02 Gew.-% Natriumazid gewaschen. Diese Röhrchen können bis zu ihrer .Verwendung bei 40C aufbewahrt werden. In jedes von drei mit Hepatitis vergesellschaftetem Antikörper überzogenen Röhrchen wird eine Plasmaprobe von lOOu 1. eingebracht. Eine dieser Plasmaproben ist die unbekannte, die anderen beiden sind negativ hinsichtlich Hepatitis vergesellschaftetem Antigen. Jede Stufe des Verfahrens wird auf jede der drei Proben angewandt. Die beiden negativen Proben ergeben die Strahlungswerte, mit denen der Strahlungswert der unbekannten Probe schließlich verglichen wird. Die Proben werden beiseite gestellt und 18 Stunden bei Raumtemperatur bebrütet. Danach werden sie mit gleichen Teilen (aliquots) des Inkubationspuffergemisches gewaschen. Zu diesem Zeitpunkt werden 2,5 ng Hepatitis vergesellschaftetem gereinigtem Antikörper mit J-125 als Tracer in 0,1 ml Volumen in jedes Röhrchen eingebracht. Die Röhrchen werden erneut beiseite gestellt und 2 Stunden .bebrütet, wonach die Röhrchen erneut mit gleichen Teilen (aliquots) an dem Inkubationspuffergemisch gewaschen werden. Jede der negativen Plasmaproben wird mit einem herkömmlichen Strahlungszähler mit einem Einsatzüchacht, der Gamma-Strahlung aufzuzeigen vermag, ausgezählt. Die negativen Proben werden 1 Minute ausgezählt, und der Mittelwert der Zählrate je Minute wird bestimmt; in diesem Fall 200 Zählungen je Minute. Dann wird die unbekannte Probe in der gleichen Weise ausgezählt, und die Zählrate wird mit dem Mittelwert je Minute der negativen Plasmaproben plus einem Korrekturfaktor gleich 50% dor Zählrate je Minute der negativen Probe verglichen. Die unbekannte Plasmaprobe hat in diesem Pail eine Zähl-
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— ι —
rate von 400 Auszählungen je Minute, die höher ist als die Zählrate 300, d.h. 200 plus $0% von 200 = 300, die das Maximum für einen negativen Test darstellt.
Das oben beschriebene Verfahren stellt einen einfachen Ja/ Nein-Test auf die Anwesenheit oder Abwesenheit von Hepatitis vergesellschaftetem Antigen in einer unbekannten Probe von Blut oder Plasma dar. Obwohl ein Korrekturfaktor erforderlich ist, läßt der Test Schlüsse zu und ist reproduzierbar und von großer Genauigkeit.
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Claims (1)

  1. 22S2413
    P ate η t a η s ρ r ji.e.h e
    [T) Verfahren zu? Feststellung der Anwesenheit von Äntigenen und ihren Antikörpern in einer unbekannten Probe, durch eine direkte Radioimmunprüfung, dadurch ge kennzeichnet , daß man
    (a) die Probe mit einem Überzug aus einem Antigen oder seinem Antikörper in Kontakt bringt;
    (b) die Probe ein erstes Mal bebrütet, während sie wiit diesem Überzug in Kontakt steht!
    (c) den bebrüteten. Überzug wäscht;
    (d) den gewaschenen Überzug mit einem einen radioaktiven Tracer enthaltenden gereinigten Antigen oder seinem Antikörper in Kontakt bringt mit der Maßgabe,, daß das den Tracer enthaltende Material das Antigen ist, wenn der Überzug das Antigen ist, und der Antikörper, wenn der Überzug der Antikörper ist\
    (e) den gewaschenen Überzug ein zweites Mal bebrütet, während er mit dem den !Radioaktiven Tracer enthaltenden Material in Kontakt steht;
    (f) den bebrüteten Überzug wäscht;
    (g) die von dem Überzug emittierte Strahlung auszählt und
    (h) die von diesem Überzug erhaltene Zählrate mit der Zählrate einer negativen, gemäß den obigen Stufen (a) bis (g) hergestellten Probe vergleicht«
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    2. Verfahren nach Anspruch 1, dadurch gekennzeichnet , daß das Antigen oder sein Antikörper
    ein Hepatitis vergesellschaftetes Antigen oder sein Antikörper ist.
    J5. Verfahren nach Anspruch 2, dadurch gekennzeichnet, daß das Hepatitis vergesellschaftete
    Antigen oder sein Antikörper als radioaktiven Tracer J-125 enthält.
    4. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß die erste Bebrütung für 1/2 bis 42 Stunden durchgeführt wird.
    5. Verfahren nach Anspruch ~5, dadurch gekennzeichnet, daß die erste Bebrütung für 1/2 bis 42 Stunden durchgeführt wird.
    6. Verfahren nach Anspruch 1, dadurch gekennzeichnet , daß das pH der unbekannten Probe mit
    einem Puffergemisch aus 0,02 Gew.-^ Natriumazid und 1 Gew.-% Rinderserumalbumen in einer 0,01m Lösung von 2-Amino-2-hydroxymethyl-ljJ-propandioL-hydrochlorid auf 6,9 bis 8,4 eingestellt wird.
    7. Verfahren nach Anspruch 3* dadurch gekennze lehnet , daß das pH der unbekannten Probe mit
    einem Puffergemisch aus 0,02 Gew.-% Natriumazid und 1 Gew.-% Rinderserumalbumen in einer 0,01m Lösung von 2-Amino-hydroxymethyl-l,j5-propandiol-hydrochlorid auf 6,9 bis 8,4 eingestellt wird.
    8. Verfahren nach Anspruch J, dadurch gekennzeichnet, daß die erste Bebrütung für 1/2 bis 42 Stunden erfolgt.
    309827/1134
    - 10 -
    ■ · ■ 41 22&2413
    9. Verfahren nach Anspruch 1, dadurch gekennz e lehnet , daß der Überzug.beide Male mit einem Puffergemisch aus 0,02 Gew.-% Natriumazid in einer 0,01m · Lösung von 2-Amino-hydroxymethyl-l,3-propandioi-hydrochloriä gewaschen wird.
    10. Verfahren nach Anspruch 8, dadurch g e'k e η η zeichnet, daß der Überzug beide Male mit einem Puffergemisch aus 0,02 Gew.-% Natriumazid in einer 0,01m Lösung von 2-Amino-hydroxymethyl-l,3-propandiol-hydrochlorid gewaschen wird.
    11. Verfahren nach Anspruch 1, dadurch gekennzeichnet-, daß die zweite. Bebrütung für 1 bis 24 Stunden bei Raumtemperatur durchgeführt wird.
    12. Verfahren nach Anspruch 10, dadurch gekennzeichnet , daß die zweite Bebrütung für 1 bis 24 Stunden bei Raumtemperatur durchgeführt wird.
    15. Verfahren zur Herstellung eines Überzuges auf einer Testvorrichtung für die direkte Radioimmunprufung auf Antigene oder deren Antikörper, dadurch gekennzeichnet, daß man
    eine Lösung eines Antigens oder seines Antikörpers in einem Puffergemisch herstellt;
    die Testvorrichtung mit dieser Lösung in Kontakt bringt;
    die Testvorrichtung 6 bis 72 Stunden bebrütet und
    die bebrütete Testvorrichtung mit dem Puffergemisch wäscht.
    lh. Verfahren nach Anspruch \J>3 dadurch gekennze ic'hnet , daß das Antigen oder sein Antikörper
    309827/1134
    - 11 -
    Hepatitis vergesellschaftetes Antigen oder sein Antikörper ist.
    15. Verfahren nach Anspruch 13» dadurch gekennzeichnet y daß das Puffergemisch 0,02 Gew.-# Natriumazid in einer 0,01m Lösung von 2-Amino-hydrcocymethyl-l,3-propandiol-hydrochlorid enthält.
    16. Verfahren nach Anspruch 14, dadurch gekennzeichnet , daß das Puffergemisch 0,02 Gew.-% Natriumazid in einer 0,01m Lösung von 2-Amino-hydroxymethyl-l,3-propandiol-hydrochlorid enthält.
    17. Verfahren nach Anspruch 16, dadurch gekennzeichnet , daß die Testvorrichtung in die Lösung getaucht wird.
    18. Verfahren nach Anspruch 17, dadurch gekennzeichnet, daß die Testvorrichtung in die Lösung
    |:j I.
    getaucht wird.
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    - 12 -
DE19722262413 1971-12-21 1972-12-20 Verfahren zur feststellung der anwesenheit eines zur bindung an zwei hepatitis-antikoerper bzw. -antigene befaehigten hepatitis-antigens bzw. -antikoerpers in einer unbekannten probe durch radioimmunpruefung Ceased DE2262413B2 (de)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US210510A US3867517A (en) 1971-12-21 1971-12-21 Direct radioimmunoassay for antigens and their antibodies

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DE2262413A1 true DE2262413A1 (de) 1973-07-05
DE2262413B2 DE2262413B2 (de) 1977-11-17

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DE19722262413 Ceased DE2262413B2 (de) 1971-12-21 1972-12-20 Verfahren zur feststellung der anwesenheit eines zur bindung an zwei hepatitis-antikoerper bzw. -antigene befaehigten hepatitis-antigens bzw. -antikoerpers in einer unbekannten probe durch radioimmunpruefung

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US (1) US3867517A (de)
AT (1) AT326818B (de)
AU (1) AU470050B2 (de)
BE (1) BE793061A (de)
CA (1) CA988420A (de)
CH (2) CH568061A5 (de)
DE (1) DE2262413B2 (de)
FR (1) FR2164705B1 (de)
GB (1) GB1414480A (de)
IT (1) IT988084B (de)
NL (1) NL7217371A (de)
PH (1) PH13655A (de)
ZA (1) ZA728798B (de)

Cited By (1)

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GB1414480A (en) 1975-11-19
FR2164705A1 (de) 1973-08-03
CH589445A5 (de) 1977-07-15
AU470050B2 (en) 1974-06-20
ZA728798B (en) 1974-02-27
AU5021472A (en) 1974-06-20
NL7217371A (de) 1973-06-25
CH568061A5 (de) 1975-10-31
FR2164705B1 (de) 1976-10-29
BE793061A (fr) 1973-06-20
US3867517A (en) 1975-02-18
PH13655A (en) 1980-08-21
ATA1088772A (de) 1975-03-15
AT326818B (de) 1975-12-29
IT988084B (it) 1975-04-10
CA988420A (en) 1976-05-04
DE2262413B2 (de) 1977-11-17

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