Antibody profiling sensitivity through increased reporter antibody layering

Abstract

A method for analyzing a biological sample by antibody profiling for identifying forensic samples or for detecting the presence of an analyte. In an embodiment of the invention, the analyte is a drug, such as marijuana, Cocaine (crystalline tropane alkaloid), methamphetamine, methyltestosterone, or mesterolone. The method comprises attaching antigens to a surface of a solid support in a preselected pattern to form an array wherein locations of the antigens are known; contacting the array with the biological sample such that a portion of antibodies in the sample reacts with and binds to the antigens in the array to form immune complexes; washing away antibodies that do form immune complexes; and detecting the immune complexes, to form an antibody profile. Forensic samples are identified by comparing a sample from an unknown source with a sample from a known source. Further, an assay, such as a test for illegal drug use, can be coupled to a test for identity such that the results of the assay can be positively correlated to the subject's identity.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation-in-part of pending U.S. patent application Ser. No. 11 / 101,254, filed Apr. 6, 2005 now U.S. Pat. No. 7,682,798, which is a divisional of U.S. patent application Ser. No. 10 / 017,577, filed Dec. 14, 2001, now U.S. Pat. No. 6,989,276, issued Jan. 24, 2006, which claims priority to U.S. Provisional Patent Application Ser. No. 60 / 290,256, filed May 10, 2001, the entire disclosure of each of which is incorporated by reference herein.CONTRACTUAL ORIGIN OF THE INVENTION[0002]This invention was made with government support under Contract No. DE-AC07-94ID13223, Contract No. DE-AC07-99ID13727, and Contract No. DE-AC07-05ID14517 awarded by the United States Department of Energy. The government has certain rights in the invention.FIELD OF THE INVENTION[0003]This invention relates to assaying biological samples. More particularly, the invention relates to methods for analyzing biological samples comprising antibody pr...

AI Technical Summary

Benefits of technology

This patent is about a method for analyzing biological material, specifically individual-specific antibodies. The method involves attaching multiple antigens to a solid support in a certain pattern, and then contacting the support with a sample of the material to form immune complexes with the antigens. The immune complexes are then detected and the pattern of their location determined, resulting in an antibody profile. The detection can be done using additional antibodies or color detection methods. The patent also describes using this method for testing drugs and comparing antibody profiles from forensic samples with those from criminals or victims of crime.

Problems solved by technology

While this method may be used to identify one individual out of a small group of individuals, the method is limited when the group of individuals is larger because no distinction is made between persons of the same blood group.

Tests based on other blood group antigens or isozymes present in body fluids suffer from the same disadvantages as the ABO blood typing tests.

These methods can exclude certain individuals, but cannot differentiate between members of the same blood group.

Although a good deal of information is known concerning the genetic makeup of the HLA locus, there are many drawbacks to using HLA serological typing for identifying individuals in a large group.

RFLP techniques are problematic, however, due to the need for relatively large sample sizes, specialized equipment, highly skilled technicians, and lengthy analysis times. For forensic applications, there is often not enough sample available for this type of assay, and in remote areas the necessary equipment is often not available.

In addition, this technique can take from two to six weeks for completion and can result in costly delays in a criminal investigation.

Moreover, the cost of RFLP analysis can be prohibitory if screening of many samples is necessary.

PCR techniques have advantages over RFLP analysis of requiring much smaller sample sizes and permitting more rapid analysis, but they still require specialized equipment and skilled technicians, and they are also expensive.

In addition, this method will potentially work on samples that contain no DNA.

While the known methods for using antibody profiling are generally suitable for their limited purposes, they possess certain inherent deficiencies that detract from their overall utility in analyzing, characterizing, and identifying biological samples.

Due to differences in conditions from one fractionation procedure to another, there are lot-to-lot differences in the positions of the antigens on the membrane such that results obtained using membranes from one lot cannot be compared with results obtained using membranes from another lot.

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