DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application is being examined under the pre-AIA first to invent provisions.
Priority
The present application was filed on 02/06/2020. This application is a CON of 12/757,685 which claims benefit of U.S. Provisional Patent Application 62/212,377 filed on 04/10/2009.
Status of the claim
Claims 117-118 are canceled. Claims 102 and 108 are withdrawn. Claims 89-101,103-107 and 109-116 are pending and examined herein.
Withdrawn Objections/Rejection
The rejection of claims 89 and 93 under 35 USC 103 as being unpatentable over Hu, Kim, Luchini and Leland is withdrawn in view of Applicant’s arguments filed on 08/29/2025.
New ground of rejection is made.
Claim Rejections - 35 USC § 103
The following is a quotation of pre-AIA 35 U.S.C. 103(a) which forms the basis for all obviousness rejections set forth in this Office action:
(a) A patent may not be obtained through the invention is not identically disclosed or described as set forth in section 102, if the differences between the subject matter sought to be patented and the prior art are such that the subject matter as a whole would have been obvious at the time the invention was made to a person having ordinary skill in the art to which said subject matter pertains. Patentability shall not be negatived by the manner in which the invention was made.
Claims 89-97, 100-101, 103 and 106-107 are rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Hu (US20080202933) in view of Boozer et al (DNA Directed Protein Immobilization on Mixed ssDNA/Oligo(ethylene glycol) Self-Assembled Monolayers for Sensitive Biosensors, Anal. Chem. 2004, 76, 6967-6972), Luchini (WO2008115653, PTO-892 09/06/2022) and Leland (US5846485; cited in IDS filed 2/6/2020).
For claims 89 and 93, Hu teaches:
A method of conducting a binding assay comprising (Hu in par.6, teaches a method including detecting an analyte bound by capture agent). Claim 93 is similar to claim 89 except the method in claim 93 is for a plurality of analytes. Hu teaches a method of conducting a binding assay for a plurality of analytes (see at least par.14, 18, 23). The method comprises
contacting (i) a sample comprising a target analyte with (ii) a particle linked to a first binding reagent that binds said target analyte, thereby forming a complex comprising said target analyte bound to said first binding reagent on the particle;
or contacting (i) a sample comprising with (ii) one or more first solid phases linked to one or more first binding reagents that bind said analytes, thereby forming complexes comprising said analytes bound to said first binding reagents on said first solid phases, (as in claim 93)
(Hu in par.5, teaching the sample is first admixed with an antibody and the antibody specifically binds to the analyte of interest; in par.9, teaching that the analyte of interest is attached to a microparticle, the microparticle is coated with at least one receptor, antibody, or anti-ligand specific for the analyte of interest.)
wherein said first binding reagent is linked to a first targeting agent and said particle is linked to a second targeting agent, and said first binding reagent and said particle are linked via a binding reaction between said first and second targeting agents, and wherein the first and second targeting agents comprise an oligonucleotide and oligonucleotide complement, respectively; (Hu in par.77-78, teaching the bound analytes can be released from the microparticles (or other solid phase) using various strategies, the binding agents having the analyte bound thereto can be freed as a complex by incorporating a cleavable or breakable bond in the linkage between the binding agents and the solid phase, the binding agent can be cleaved using proteases or nucleases (e.g., sequence-specific proteases or nucleases), nucleic acid binding molecule can be cleaved with nucleases.
It appears that the linkage encompasses a complex of a first and a second targeting agent, so that the antibody can be immobilized on the solid phase. Also, Hu gave an example that the nucleic acid binding molecule can be a linker in par.78 so that it can be cleaved with nucleases. The nucleic acid binding molecule encompasses oligonucleotide. Hu does not clearly teach using oligonucleotide and oligonucleotide complement for a linkage.)
(b) collecting said complex on said particle in a first assay volume;(c) washing said complex on said particle to remove unbound components of said sample from said first assay volume; (Hu in par.16, teaching microparticles to which primary antibodies are bound are used as a solid phase to bind analyte in a large volume reservoir (i.e., first assay volume). Hu in par.73, teaching after attachment of the analyte to the microparticles, an analyte-enriched fraction can be prepared by segregating the microparticles using centrifugation, magnetic separation, or filtration, with appropriate washing steps; removal of unbound contaminants using other solid phases (e.g., microplate wells) can be accomplished by removing the unbound supernatant, and other well-known methods)
(d) disposing said washed complex on said particle in a second assay volume, wherein said second assay volume is smaller than said first assay volume; (e) releasing said complex from said particle in the second assay volume by disassociating said binding interaction between said first and second targeting agents, thereby yielding a concentrated sample of said complex; (Hu in par.26 and 30, and Fig.1: showing that the large volume reservoir (LVR) (i.e., first assay volume) is fluidically connected to a staging reservoir (i.e., second assay volume). Hu in par.126, teaching the bound analyte/antibody complex after washed is then electrophoresed to concentrate the analyte 3 into a staging reservoir. Hu in par.89: teaching that the volume of the staging reservoir is smaller than the volume of the large volume reservoir.
Hu in par.126, teaching the bound analyte/antibody complex is then removed from the microparticle using an appropriate cleaving agent. Hu in par.35 teaching analyte is being concentrated or accumulating in the staging reservoir.
While Hu’s method at the end still get to yield a concentrated sample of said complex in the second assay volume by dissociating the binding interaction between the first and second targeting agents, Hu teaches that the releasing process of the washed complex from particle happens in the first assay volume, not in the second assay volume.)
(f) contacting said concentrated sample with a solid phase linked to a second binding reagent that binds to said complex or a component thereof to form a solid phase-bound complex, (Hu in par.6 teaches that the analysis area is a capture site including at least one capture agent and this allows the analyte of interest to be captured. Hu in par.101 teaches that the capture agent can be immobilized to the microfluidic device at a capture site. Hu in par.100 discloses that the capture agent is a molecule that can capture an analyte or analyte complex by specifically binding the analyte or a member of the complex.)
wherein the solid phase comprises an electrode, wherein the second binding reagent is an antibody; (Hu in par.9 teaches a second antibody can be provided and can bind to a different site on the analyte. Hu in par.100 teaches the capture agent can be, for example, one or more antibodies. Hu in par.114 teaches that the solid phase comprises an electrode associated with the analysis area.)
(g) applying a voltage waveform to said electrode; (Hu in par.33, teaching that the electrodes can be attached or positioned in any way that allows an electric field for electrophoresis to be generated when the current is on and solution is present to complete a circuit; in par.93 teaching voltages to be applied in such a way that analytes can be transferred with speed and minimizing the voltage required to transport analytes into the microfluidic chip)
and (h) measuring signal label attached to the first binding reagent and correlating said signal with an amount of analyte in said sample. Hu in par.105 teaches analyte is detected based on a signal from a detectable label, wherein detectable labels suitable for use in the present invention include any composition detectable by spectroscopic, photochemical, biochemical, immunochemical, electrical, optical, chemical or physical means and the like. Hu in par.126 teaches a detectable label 7 can be attached to any of the molecules involved, including the analyte 3, the antibody binding molecule 11 on the microparticle 8, the second antibody 1, or the ionic moiety.
Hu does not clearly teach using oligonucleotide and oligonucleotide complement for a linkage. Hu does not teach that the releasing process of the washed complex from particle happens in the second assay volume. Hu does not clearly teach part (h) which measuring an electro-chemiluminescent (ECL) signal emitted by an ECL.
Boozer teaches a technique for immobilizing a protein or an antibody on a biosensor surface such as nanoparticles (see Abstract, see page 6967 col.2 par.1). Boozer teaches a surface linked to a first binding reagent wherein said first binding reagent is linked to a first targeting agent (see page 6967 column 2 par.2: simple DNA-antibody conjugate) and said particle is linked to a second targeting agent (see page 6967 column 2 par.2: a single-stranded DNA (ssDNA) probe surface). Boozer teaches said first binding reagent and said particle are linked via a binding reaction between said first and second targeting agents (see page 6967 col.2 par.2: the protein conjugate consists of an antibody chemically linked to a ssDNA target with a sequence complementary to the surface-bound ssDNA probe; DNA hybridization will result in immobilization of the antibody on the surface).
Boozer teaches the DNA in DNA-antibody conjugate is oligonucleotide probes (see page 6968 col.2: the surface is patterned with the appropriate complementary oligonucleotide probes, a mixture of the conjugates can be applied to the surface and sequence-specific hybridization will direct the target conjugates to the appropriate spots on the surface).
Boozer teaches biosensor surface can be regenerated because the DNA double helix can be de-hybridized, or melted, by either chemical or thermal means (see page 6971 col.1 par.2).
Luchini discloses a microfluidic system for detecting an analyte in a sample comprising collecting the analyte of interest on a capture particle in the first location, transferring the analyte bound capture particles to the second location where the analyte is released and analyzed using methods known in the art (see par.15-16). Luchini teaches releasing analyte in a second assay volume, thereby releasing analyte from a particle to yield a concentrated sample, wherein the second assay volume is smaller than the first assay volume (see par.59). Luchini teaches this allows analytes having concentrations below the limits of detection of various analytical methods in their original fluids to be concentrated so that they have a concentration that can easily be detected and analyzed (par.59).
Leland in col.18, ln.13-44, teaches that electro-chemiluminescent (“ECL) assay can apply to detect or quantify of an analyte of interest in a given sample. Leland in col.19, ln.4-9, teaches it should be possible to coat the surface of an electrode with antibodies, so that only antigen linked directly or through one or more other molecules to the ECL moiety and bound to the immobilized antibodies can obtain access to the electrode and thereby be determined. Leland teaches applying a voltage waveform to the electrode (see col.14 ln.60-67 and col.15 ln. 1-7, teaching that ECL reactive mixture is triggered to emit light by a voltage impressed on the working electrode). Leland teaches measuring (ECL) signal emitted by an ECL label linked to the complex or a component thereof and correlating the signal with an amount of analyte in the sample (see col.13, ln.9-46).
Therefore, it have been obvious to one having ordinary skill in the art before the effective filing date of the claimed invention to modify the linkage of Hu’s method in the part (a) with an oligonucleotide and its complement as taught by Boozer because Hu is generic for the bonds of the linkage as long as they can be dissociated by chemicals or enzymes, thereby the bound analytes can be released from the microparticles (or other solid phase) (see Hu par.77-78) and Hu provides a nucleic acid binding agent as an example for the linkage (see par.78), which encompasses an oligonucleotide agent. Booze supports for the use of oligonucleotide as a linker for attaching antibody on a particle surface. One would have been motivated to use the technique taught by Booze because this protein-resistant oligonucleotide background chip is crucial for the proper placement of the antibody conjugates and minimizes the chance of false detections caused by nonspecific adsorption (see page 6972 col.1 par.2). The chip is more stable than a standard protein chip and can be regenerated (Booze page 6968 col.2 par.1).
The combination would have yield predictable result because Booze teaches the oligonucleotide background chip can be regenerated because DNA hybridization is reversible (see page 6968 col.2 par.1), which fits the requirement of Hu for the linkage on the analyte-captured microparticle.
It would have been obvious to one having ordinary skill in the art before the effective filing date of the claimed invention to modify the releasing step of Hu such that the second assay volume is smaller than the first assay volume before releasing the first binding reagent from the particle yields a concentrated sample of analyte bound to the first binding reagent in the complex as taught by Luchini in order to allow analytes having concentrations below the limits of detection of various analytical methods in their original fluids to be concentrated so that they have a concentration that can easily be detected and analyzed (Luchini, par. 59). It is because the method of Luchini and Hu are functional equivalent. While Hu does not teach the releasing process happening in the second assay volume, Hu’s method at the end still get to yield a concentrated sample of said complex in the second assay volume by dissociating the binding interaction between the first and second targeting agents (see at least par.35 and 90: teaching that the analyte is being concentrated or accumulating in the staging reservoir).
Since Applicant has not disclosed that the specific limitation (i.e., dissociating the analyte-antibody complex from the particle) recited in the second assay volume is for any particular purpose other than concentrating sample of said complex in the second assay volume (see at least in instant specification page 19 last paragraph), therefore, the claimed limitation is not sufficient by itself to patentably distinguish over the teaching of Hu or Luchini because it is directed to dissociate the binding interaction between the first and second targeting agents to concentrate the analyte of interest before being analyzed. This step is essential for measuring the analyte in the sample that has analyte concentrations below the limits of detection of various analytical methods in their original fluids. Thus, it would be an obvious matter of design choice to concentrate the target analyte before or after transferring the target analyte to the second volume (as taught by Hu or Luchini respectively) since the changes in sequence of adding ingredients, the separation and/or rearrangement of the method steps are all directed to yield a concentrated analyte before going to analysis step (see MPEP 2144.04 (IV)(C), (V)(C), and (VI)(C)).
One having ordinary skills in the art would have had a reasonable expectation of success in combining the prior art references because Hu and Luchini are similarly drawn to methods of using a particle to isolate analyte for analysis, concentrating the analyte from the original sample before measuring the present of analyte.
It would have been obvious to one having ordinary skill in the art before the effective filing date of the claimed invention to substitute a label attached to the first binding reagent in Hu by an ECL label and use an electrode for the solid phase or additional solid phase of Hu to measure a signal emitted by a ECL label as taught by Leland because Leland teaches the emission of ECL signal correlates with an amount of analyte in the sample, thus means the final goal ECL measurement is also measuring or quantitating the concentration of an analyte in the sample. Since Hu is generic with respect to the label, and the analysis steps of the complex, one would be motivated to use the appropriate solid phase, label, and analysis techniques for detection of the desired analyte. Moreover, Leland teaches using ECL to provide detection and quantitation of an analyte of interest over a wide concentration range, down to a very small analyte concentration, in aqueous as well as organic environments, for example, without disrupting the immunochemistry of highly interesting biological systems which exist at physiological pH.
One having ordinary skills in the art would have had a reasonable expectation of success in combining the prior art references because Hu is generic with respect to how the analyte is detected or analyzed and the microchip taught by Hu is totally appropriate for using ECL detection because the chip has one or more electrodes that can be applied a voltage waveform on as described in Leland. Thus it is obvious that the Hu method can be incorporated into the method of Leland if a ECL label is used.
For claims 90 and 94, Hu, Boozer, Luchini and Leland teach the method of claim 89 or 93, wherein said releasing step comprises subjecting said complex to increased temperature, pH changes, nucleic acid denaturant, and combinations thereof.
(Hu in par.77 teaches the bound analytes can be released from the microparticles (or other solid phase) using various strategies, e.g., denaturing agents, extreme pH, temperature. This can be done using changes in buffer, changes in the electric field, pH, addition of an elution buffer, or any method known in the art)
For claims 91 and 95, Hu, Boozer, Luchini and Leland teach the method of claim 89 or 93, wherein said collecting step comprises a method selected from the group consisting of centrifugation, gravity, filtration, magnetic collection, and combinations thereof.
(Hu in par.73 teaches the collecting step comprises centrifugation, filtration, or magnetic collection).
For claims 92 and 96, Hu, Boozer, Luchini and Leland teach the method of claim 89 or 93, wherein said assay is a sandwich assay or a competitive assay.
(Hu in Fig. 5-6, par. 126-128 teaches a sandwich assay, Hu in par.77 teaches an analyte can be displaced using specific agents to compete with the analyte for binding to the immobilized SBP, thus it is a competitive assay).
For claims 97 and 103, Hu, Boozer, Luchini and Leland teach the method of claim 89 or 93, wherein the first and second binding reagents each comprise antibodies (Hu in par.9 teaches the microparticle is coated with antibody, a second antibody can be provided and can bind to a different site on the analyte, see more in par.126).
For claims 100-101 and 106-107, Hu, Boozer, Luchini and Leland teach the invention methods of claims 89 and 93 as discussed above. Hu teaches the solid phase or additional solid phase is located within an assay device, such as an assay chip (Hu in par.78 teaches a microfluidic chip).
Hu teaches the solid phase is located within a well of a multi-well assay plate (see par.71).
For claims 98-99 and 104-105, Hu, Boozer, Luchini and Leland teach the invention methods of claims 89 and 93 as discussed above. Hu does not teach the length of the oligonucleotide and its complement comprising 10 to 50 bases, or 10 to 25 bases.
Boozer further teaches the length of the oligonucleotide and its complement comprising 10 to 50 bases, or 10 to 25 bases (see page 6971 col.1 par.1: the 24-base pair-long DNA used in this work acts like a molecular tether and provides the immobilized antibody with greater mobility than a biotin/streptavidin linkage).
Therefore, it have been obvious to one having ordinary skill in the art before the effective filing date of the claimed invention to modify the linkage of Hu’s method in the part (a) with an oligonucleotide and its complement as taught by Boozer, with the length about 24-base pair-long because it provides the immobilized antibody with greater mobility than a biotin/streptavidin linkage so that the oligonucleotide background chip obtains the superior sensitivity (see page 6970 col.2 par.3). One would have been motivated to use the technique taught by Booze because this protein-resistant oligonucleotide background chip is crucial for the proper placement of the antibody conjugates and minimizes the chance of false detections caused by nonspecific adsorption (see page 6972 col.1 par.2). The chip is more stable than a standard protein chip and can be regenerated (Booze page 6968 col.2 par.1).
The combination would have yield predictable result because the combined teaching provides the elements as claimed by known methods from Hu and Booze with no change in their respective functions.
Claims 109-116 are rejected under pre-AIA 35 U.S.C. 103(a) as being unpatentable over Hu in view of Boozer, Luchini and Leland, as applied to claims 89 and 93 above, and further in view of Nadeau (US 2005/0009050 A1; cited in IDS filed 2/6/2020).
Regarding claims 109-116, Hu, Boozer, Luchini and Leland teach the invention methods of claims 89 and 93 as discussed above. They do not teach the first targeting agent oligonucleotide sequence is longer than the second targeting agent oligonucleotide sequence.
Nadeau discloses methods for attaching antibodies to solid surfaces such as a particle via oligonucleotide hybridization (see par.119). Nadeau in par.119, par.125 and Fig. 6E, teaches the antibody is conjugated to an oligonucleotide P1 via its 5′ terminus, the conjugated oligonucleotide comprises a sequence that is complementary to another oligonucleotide P2 that is attached by its 5′ terminus to a solid surface.
Nadeau in par.154-162 provides examples of P1 and P2 sequences. For example:
P1 in par.155: 5' CCA GTC TTG TCT TGT CTG TTC TCG GGA TGC ATT CAG TGA CGT GAT GAG CTA GAC AGA TGT ACA GT,
P2 in par.158: 5' ATT CAC GCT TCC ATT СCA TGT CTC GGG TTT ACT TCA TCT GCA ACT GTA C,
which P1 is longer than P2 16 bases.
P2 in par.159: 5' ATT CAC GCT TCC ATT CCA TGT CTC GGG TTT ACT ТCA TCT GCA ACT GTA CAT,
which P1 is longer than P2 14 bases.
P2 in par.160: 5' ATT CAC GCT TCC ATT CCA TGT CTC GGG TTT AСТ ТСА ТCT GCA ACT GTA CAT CTGT,
which P1 is longer than P2 10 bases.
Therefore, it have been obvious to one having ordinary skill in the art before the effective filing date of the claimed invention to substitute the linker of the modified method of Hu, using the linker taught by Nadeau which having the first oligonucleotide sequence is longer than the second oligonucleotide sequence (can be longer up to 25 or 15 or 10 bases), for immobilizing an analyte-specific antibody to a particle because Nadeau supports that the oligonucleotide sequences in pair can immobilize an antibody on a solid surface, thus the linkers are functionally equivalent. Moreover, Nadeau also teaches doing that would provide a high-sensitivity, low-background immuno-amplification assay (see Abstract).
One of ordinary skill in the art could have combined the elements as claimed by known methods from Hu and Nadeau with no change in their respective functions, and the combination yielded nothing more than predictable results because the combination teaches each and every claimed elements for attach antibody on a solid surface: a particle linked to a first binding reagent that binds said target analyte (e.g., particle links to antibody), wherein said first binding reagent is linked to a first targeting agent (antibody links to oligonucleotide P1) and said particle is linked to a second targeting agent (particle links to oligonucleotide P2), and said first binding reagent and said particle are linked via a binding reaction between said first and second targeting agents (P1 hybridizes to P2), and wherein oligonucleotide sequence P1 is longer than oligonucleotide sequence P2 (up to 25 bases longer, or up to 15 bases longer, or up to 10 bases longer) as discussed above.
Response to Arguments
The Applicant's arguments filed on 08/29/2025 have been fully considered but moot because new ground of rejection is made.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHAU N.B. TRAN whose telephone number is (571)272-3663. The examiner can normally be reached Mon-Fri 8:30-6:30 CT.
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/CHAU N.B. TRAN/Examiner, Art Unit 1677
/BAO-THUY L NGUYEN/Supervisory Patent Examiner, Art Unit 1677