Haptenic probes for detecting capture polynucleotides

• US 5,552,541 A
• Filed: 06/20/1990
• Issued: 09/03/1996
• Est. Priority Date: 06/20/1990
• Status: Expired due to Term

• Current Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Original Assignee: Beckman Instruments, Inc. (Danaher Corporation)
• Inventors: Adams, Craig W.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): Marschel, Ardin H.
• Application Number:
  US07/541,143
• Time in Patent Office: 2,267 Days
• Field of Search: 435/6, 435/7.1, 435/7.5, 435/7.7, 435/7.92, 436/501, 536/27, 536/22.1, 536/23.1, 536/24.1, 536/24.3-.33, 536/25.3
• US Class Current: 536/22.1
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/125 the label being enzymatic, i.e. proteins, and non proteins, such as nucleic acid with enzymatic activity
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence

Abstract

Nucleic acid probes and protein probes are disclosed. The nucleic acid probe comprises a probe polynucleotide, a charged hapten label, and a binding moiety. The protein probe comprises a probe protein, a charged hapten label and a binding moiety. The charged hapten label joint to the binding moiety can comprise a negatively charged sulfophenylhydrazine tag compound. Polyclonal antibodies and monoclonal antibodies with specific affinity for the charged hapten labels are disclosed as are hybridomas capable of making the monoclonal antibodies. Methods and kits are disclosed for making the nucleic acid probes, making the protein probes, detecting a capture polynucleotide of the nucleic acid probe and detecting a capture molecule of the protein probe.

Claims

1. A non-radioactive, haptenic probe for detecting a capture polynucleotide comprising:

(a) a probe polynucleotide, the probe polynucleotide having a polynucleotide sequence substantially complementary to a polynucleotide sequence of the capture polynucleotide;

(b) at least one non-radioactive, charged hapten label selected from the group consisting of benzoic acid anion, 4-(sulfonyl) benzoic acid anion, and sulfophenyl anion; and

(c) a hydrazine binding moiety covalently attaching the charged hapten to the N-4 position of a cytosine base of a nucleotide located on the probe polynucleotide wherein detection of the capture polynucleotide is based upon an immunological reaction between the non-radioactive, charged hapten label and an antibody specific to the label.

2. The probe of claim 1 further comprising a spacer compound between the binding moiety and the charged hapten label for separating the charged hapten label from the probe polynucleotide, the spacer compound comprising a linear chain of from about two to about fifteen carbon atoms.

3. The probe of claim 1 wherein the probe polynucleotide comprises single-stranded RNA.

4. The probe of claim 1 wherein the hapten label and the binding moiety comprise a sulfophenylhydrazine compound.

5. The probe of claim 4 wherein the sulfophenylhydrazine compound comprises a 4-hydrazinobenzenesulfonate anion.

6. A hybridization probe comprising a single-stranded DNA or RNA polynucleotide comprising a cytosine base, wherein a 4-hydrazinobenzenesulfonate anion is covalently attached through the hydrazide terminus thereof to the 4-position of the cytosine base.

7. A hybridization probe capable of hybridizing with a capture polynucleotide, the hybridization probe comprising a 4-hydrazinobenzenesulfonate anion covalently attached through its hydrazine terminus to the 4-position of a cytosine base of the probe polynucleotide.