Method for simultaneous quantitative analysis of several constituents in a sample

• US 4,094,759 A
• Filed: 01/12/1977
• Issued: 06/13/1978
• Est. Priority Date: 01/03/1975
• Status: Expired due to Term

• Current Assignee: Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften e.V. (Max-Planck-Gesellschaft zur Frderung der Wissenschaften e)
• Original Assignee: Max-Planck-Gesellschaft Zur Foerderung Der Wissenschaften e.V. (Max-Planck-Gesellschaft zur Frderung der Wissenschaften e)
• Inventors: Ruhenstroth-Bauer, Gerhard, Scherer, Reiner
• Primary Examiner(s): Prescott, Arthur C.
• Application Number:
  US05/758,735
• Time in Patent Office: 517 Days
• Field of Search: 204/180 S, 204/180 G, 204/299
• US Class Current: 436/516
• CPC Class Codes: G01N 27/447 using electrophoresis
• CPC Class Codes: G01N 33/561 Immunoelectrophoresis

Abstract

A method for simultaneous qualitative and quantitative analysis of antigensr antigen-effective substances in a sample provides placing the sample in a carrier medium through which it migrates toward a plurality of physically separate carrier strips. The medium on each strip contains a quantity of a specific ingredient, for example an antibody, which is expected to undergo a specific immunoreaction with one of the constituents in the sample, thereby producing a visible precipitate. The dimensions of the final precipitate then permit a direct measurement of the identified substance. A known concentration of a calibrating substance may be added to the sample to provide for calibration precipitates in the various strips. The apparatus for practicing the method includes a base plate on which the neutral, antibody-free carrier medium as well as the various strips containing media with antibodies or other immunoreactive ingredients are located. An electric field may be applied to aid in the migration of the sample in the medium.

Claims

1. A method for the simultaneous qualitative and quantitative analysis of a plurality of immunoreactive substances, especially of antigen-effective substances contained in a sample, comprising the steps of:

A. placing the sample to be tested in a first carrier material which is free from antibodies;

B. providing a second carrier material, adjacent to said first carrier material, said second carrier material containing ingredients capable of undergoing immunoreactions with said substances in said sample, thereby forming precipitates;

C. causing the migration of said sample from said first carrier material into said second carrier material;

D. applying an electric field to the area of said first and second carriers for enhancing the migration of said sample from said first carrier material into said second carrier material substantially in the direction of said electric field;

the improvement comprising the steps of;

E. providing a plurality of second carrier materials, simultaneously adjacent to said first carrier material, and

F. providing in each of said plurality of second carrier materials a different ingredient, each different ingredient being capable of undergoing a precipitate-forming immunoreaction with one of the substances to be analyzed in said sample.

2. A method as defined in claim 1, further comprising the step of:

G. treating the sample to alter the chemical electric charge on the molecules in said sample so as to obtain a negative effective charge.

3. A method as defined in claim 2, wherein the sample is treated with a chemical from the group comprising α

and β

-propiolactene and potassium cyanate.

4. A method as defined in claim 2, wherein the sample is treated for approximately 30 minutes at approximately 20°

Centigrade.

5. A method as defined in claim 1, wherein the step of applying an electric field includes applying an electric field of approximately 8 volts centimeter for approximately 8-10 hours.

6. A method as defined in claim 1, further comprising the steps of:

H. adding to the sample a known concentration of a known calibrating substance different from any substance to be analyzed;

I. adding to each of said second carrier materials a calibrating ingredient different from said ingredients which undergo immunoreaction with said substances to be analyzed, said calibrating ingredient being capable of undergoing an immunoreaction with said calibrating substance.

7. A method as defined in claim 1, wherein the concentration of ingredients in said plurality of second carrier materials is so chosen that, when a sample with a predetermined concentration of antigen -effective substances is placed in said first carrier material, the immunoprecipitates formed in each of said plurality of second carrier materials all have substantially the same geometrical extent, as measured from the limits of said first carrier material.

8. A method as defined in claim 1, wherein said antigen-effective substances are antigens.

9. A method as defined in claim 1, wherein said sample is blood plasma and said antigen-effective substances are plasma proteins.

10. A method as defined in claim 1, wherein said sample is blood serum and said ingredients in said second carrier materials are different tissue extracts;

whereby the resulting precipitates are indicators of the phenomenon of auto-immunoreaction.