公开(公告)号：US4642285A

公开(公告)日：1987-02-10

申请号：US606122

申请日：1984-05-02

申请人： Seymour P. Halbert ,  Milton Anken

发明人： Seymour P. Halbert ,  Milton Anken

IPC分类号： G01N33/576 ,  G01N33/53 ,  G01N33/535 ,  G01N33/545

CPC分类号： G01N33/576 ,  Y10S435/805 ,  Y10S435/81 ,  Y10S436/811 ,  Y10S436/82

摘要： A sensitive direct immunoassay system is provided for the detection of an antigen in body fluids. A single antibody which reacts with an antigen or antigens and which is bonded to an insoluble member, is incubated with a test sample. During this first period of incubation a portion of an antigen present in the test sample will combine with the antibody immobilized on the insoluble member. The antibody bonded member, to which antigen is attached, is then washed and incubated with an enzyme tagged antibody reagent. During the second incubation, the tagged antibody reacts with antigen fixed to the antibody member in the first incubation. Thus, an immobilized "sandwich" is formed of an insoluble member- antibody-antigen-enzyme tagged antibody. After the second incubation, the member is washed again to remove unreacted enzyme antibody reagent. The member is then exposed to a substrate which is converted by the enzyme to produce an end product. The tagged antibody reagent will be fixed in the second incubation only if antigen was present in the sample. The amount of enzyme tagged antitbody fixed is proportional to the amount of antigen or antigens present in the test sample up to the maximum capacity of the test. The concentration of the end product, and hence the amount of antigen or antigens, is determined by a spectrophotometer which measures the optical absorption of light by the end product. This readout is then compared against a standard value for both antigen negative and antigen positive samples.

摘要翻译： 提供敏感的直接免疫测定系统用于检测体液中的抗原。 与抗原或抗原反应并与不溶性成分结合的单一抗体与测试样品一起温育。 在该第一次孵育期间，存在于测试样品中的部分抗原将与固定在不溶性成分上的抗体结合。 然后将抗体连接的抗体结合成员洗涤并与酶标记的抗体试剂一起温育。 在第二次孵育期间，标记的抗体在第一次孵育中与固定在抗体成员上的抗原反应。 因此，固定的“夹心”由不溶性成员 - 抗体 - 抗原 - 酶标记的抗体形成。 第二次孵育后，再次洗涤成员以除去未反应的酶抗体试剂。 然后将该构件暴露于由酶转化以产生最终产物的底物。 只有抗原存在于样品中时，标记的抗体试剂才能在第二次孵育中固定。 固定的酶标抗体的量与测试样品中存在的抗原或抗原的量成比例，直到测试的最大容量。 最终产物的浓度，因此抗原或抗原的量由分光光度计测定，该分光光度计测量最终产物对光的光吸收。 然后将该读数与抗原阴性和抗原阳性样品的标准值进行比较。

公开(公告)号：US4157280A

公开(公告)日：1979-06-05

申请号：US617745

申请日：1975-09-29

申请人： Seymour P. Halbert ,  Milton Anken

发明人： Seymour P. Halbert ,  Milton Anken

IPC分类号： G01N33/545 ,  G01N33/576 ,  G01N31/14 ,  G01N33/16

CPC分类号： G01N33/576 ,  G01N33/545 ,  Y10S436/82

摘要： A test set useful for detecting the presence of antigens associated with hepatitis comprising an insoluble, polymeric disc-like solid having antibody reactive with antigens associated with hepatitis bonded thereto, an enzyme-hepatitis antibody conjugate reagent, an enzyme substrate solution, control sera negative, weakly positive, and strongly positive for antigens associated with hepatitis, an enzyme substrate buffer, and a horse globulin solution. Preferably, the enzyme of the conjugate is alkaline phosphatase, the enzyme substrate is p-nitrophenylphosphate and the substrate buffer has a pH on the order of 9.8.

摘要翻译： 一种用于检测与肝炎相关的抗原存在的测试集，其包含不溶性聚合物盘状固体，其具有与与其结合的肝炎相关的抗原反应的抗体，酶 - 肝炎抗体缀合试剂，酶底物溶液，对照血清阴性， 弱阳性，与肝炎相关抗原，酶底物缓冲液和马球蛋白溶液强阳性。 优选地，缀合物的酶是碱性磷酸酶，酶底物是对硝基苯基磷酸盐，底物缓冲液的pH值为9.8。

公开(公告)号：US4317810A

公开(公告)日：1982-03-02

申请号：US126525

申请日：1980-03-02

申请人： Seymour P. Halbert ,  Milton Anken

发明人： Seymour P. Halbert ,  Milton Anken

IPC分类号： G01N33/545 ,  G01N31/06 ,  G01N33/48 ,  G01N33/52 ,  G01N33/54

CPC分类号： G01N33/545 ,  Y10S436/809 ,  Y10S530/816

摘要： A disc-shaped insoluble matrix is formed of a polymeric material having a layer of reactive groups which react with proteins binding them covalently and uniformly to its surface. An antibody adapted for use in immunoassay of antigens associated with hepatitis is covalently bonded to the reactive groups. The matrix is provided with irregular, e.g., waffle-like, surfaces designed to increase the amount of antibody that will be in contact with a test solution such as serum.

摘要翻译： 盘状不溶性基质由具有反应性基团的聚合物材料形成，所述反应性基团与与其表面共价均匀结合的蛋白质反应。 适用于免疫测定与肝炎相关的抗原的抗体与反应性基团共价键合。 该基质具有不规则的，例如华夫饼样的表面，其设计用于增加将与诸如血清的测试溶液接触的抗体的量。

公开(公告)号：US4474878A

公开(公告)日：1984-10-02

申请号：US617743

申请日：1975-09-29

申请人： Seymour P. Halbert ,  Milton Anken

发明人： Seymour P. Halbert ,  Milton Anken

IPC分类号： G01N33/545 ,  G01N33/576 ,  G01N33/54 ,  G01N33/58

CPC分类号： G01N33/545 ,  G01N33/576 ,  Y10S435/805 ,  Y10S435/81 ,  Y10S436/811 ,  Y10S436/82

摘要： A sensitive direct immunoassay system is provided for the detection of an antigen associated with hepatitis in body fluids. A single antibody which reacts with a hepatitis antigen or antigens and which is bonded to an insoluble member, is incubated with a test sample. During this first period of incubation a portion of an antigen present in the test sample will combine with the antibody immobilized on the insoluble member. The antibody bonded member, to which antigen is attached, is then washed and incubated with an enzyme tagged antibody reagent. During the second incubation, the tagged antibody reacts with antigen fixed to the antibody member in the first incubation. Thus, an immobilized "sandwich" is formed of an insoluble member- antibody-antigen-enzyme tagged antibody. After the second incubation, the member is washed again to remove unreacted enzyme antibody reagent. The member is then exposed to a substrated which is converted by the enzyme to produce an end product. The tagged antibody reagent will be fixed in the second incubation only if antigen was present in the sample. The amount of enzyme tagged antibody fixed is proportional to the amount of antigen of antigens present in the test sample up to the maximum capacity of the test. The concentration of the end product, and hence the amount of antigen or antigens, is determined by a spectrophotometer which measures the optical absorption of light by the end product. This readout is then compared against a standard value for both antigen negative and antigen positive samples.

摘要翻译： 提供敏感的直接免疫测定系统用于检测体液中与肝炎相关的抗原。 将与抗原或抗原反应并与不溶性成分结合的单一抗体与测试样品一起温育。 在该第一次孵育期间，存在于测试样品中的部分抗原将与固定在不溶性成分上的抗体结合。 然后将抗体连接的抗体结合成员洗涤并与酶标记的抗体试剂一起温育。 在第二次孵育期间，标记的抗体在第一次孵育中与固定在抗体成员上的抗原反应。 因此，固定的“夹心”由不溶性成员 - 抗体 - 抗原 - 酶标记的抗体形成。 第二次孵育后，再次洗涤成员以除去未反应的酶抗体试剂。 然后将该成员暴露于由酶转化以产生最终产物的底物。 只有抗原存在于样品中时，标记的抗体试剂才能在第二次孵育中固定。 固定的酶标记抗体的量与测试样品中存在的抗原的抗原量成比例，直到测试的最大容量。 最终产物的浓度，因此抗原或抗原的量由分光光度计测定，该分光光度计测量最终产物对光的光吸收。 然后将该读数与抗原阴性和抗原阳性样品的标准值进行比较。