DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Claims
The Amendment filed October 21st, 2025 has been entered. Claims 1-2, 6, 17-18, 22, 26, and 29 have been amended. Claims 31-32, 34, and 47-48 have been previously withdrawn. Claim 10 has been canceled with Claims 5, 7-19, 11-14, 16, 19-20, 23, 27-28, 30, 33, 35-46, and 49-58 previously canceled. Claim 59 has been added. Claims 1-4, 6, 15, 17-18, 21-22, 24-26, 29, and 59 are currently examined herein.
Status of the Rejection
Applicant’s amendments to the claims have overcome each objection and 112(b) rejection previously set forth in the Non-Final Office Action mailed April 22nd, 2025.
All 35 U.S.C. § 102 and 35 U.S.C. § 103 rejections from the previous office action are withdrawn in view of Applicant’s amendments.
New grounds of claim objection and rejection under 35 U.S.C. § 103 are necessitated by the Applicant’s amendments as outlined below.
Claim Objections
Claim 1 is objected to because of the following informalities:
Claim 1, please amend “the microfluidic device maintained at a temperature…” to “the microfluidic device is maintained at a temperature…”.
Appropriate correction is required.
Claim Rejections - 35 USC § 103
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-4, 6, 15, 21-22, 24, and 29 is rejected under 35 U.S.C. 103 as being unpatentable over Thanthri (Simultaneous Preconcentration and Separation of Native Protein Variants Using Thermal Gel Electrophoresis,” Analytical Chemistry, Vol. 92, 2020, pages 6741-6747, provided in IDS submitted on 08/15/2023) in view of Liu (Optimizing capillary gel electrophoretic separations of oligonucleotides in liquid crystalline Pluronic F127. Journal of Chromatography A. 1998; 817, pages 367-375).
Regarding Claim 1, Thanthri teaches a method of injectionless gel electrophoresis (thermal gel electrophoresis [TGE] used for separation of protein variants [para. 2 col. 1, page 6742]; no injection step was used [third para. col. 1, page 6743]), comprising:
loading a mixed analyte sample (Ovalbumin-Texas Red dye conjugate [third para. col. 1 [page 6742]; ovalbumin protein sample was a mixed analyte sample containing multiple ovalbumin protein variants that were separated [second para. col. 2, page 6743; separation illustrated in Figure 2B]) mixed with a gel solution (ovalbumin mixed with PF-127 [third para. col. 1, page 6743]) into a channel of a microfluidic device (PF-127 and protein sample directly cast into a single-channel microfluidic device [third para. col. 1, page 6743]; microfluidic device described in [second para. col. 2, page 6742]), the channel having a first end (as illustrated in Figure 1, single-channel microfluidic device has a first end on the left side [page 6742]) and a second end (as illustrated in Figure 1, single-channel microfluidic device has a second end on the right side [page 6742]), the microfluidic device having a first reservoir coupled to the first end of the channel (as illustrated in Figure 1, a first reservoir containing trailing electrolyte [TE] located on left side of the single-channel microfluidic device and is coupled to the first end of the channel [page 6742]) and a second reservoir coupled to the second end of the channel (as illustrated in Figure 1, a second reservoir containing leading electrolyte [LE] located on right side of the single-channel microfluidic device is coupled to the second end of the channel [page 6742]);
providing a first reservoir solution in the first reservoir (trailing electrolyte [TE] provided as a first reservoir solution in the first reservoir [third para. col. 2, page 6742]; illustrated in Figure 1 [page 6742]);
providing a second reservoir solution in the second reservoir (leading electrolyte [LE] provided as a second reservoir solution in the second reservoir [third para. col. 2, page 6742]; illustrated in Figure 1 [page 6742]); and
applying an electric field across the microfluidic device (electric field applied across microfluidic channel [first para. col. 2, page 6742]).
Thanthri is silent on the microfluidic device is maintained at a temperature between 45⁰C and 60⁰C.
Liu teaches gel electrophoresis separations at various temperatures for separating oligonucleotides (abstract), and although Liu does not teach the microfluidic device maintained at a temperature between 45⁰C and 60⁰C, Liu teaches the microfluidic device maintained at a temperature between 20⁰C and 50⁰C (oligonucleotides are separated in 20% Pluronic gels from 20⁰C and 50⁰C [col. 2 second and third paras., page 370; CGE separations with respect to temperature shown in Figure 4, page 371]).
Thanthri and Liu are considered analogous art to the claimed inventions because they are in the same field of methods of electrophoresis. It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the temperature of the microfluidic device of Thanthri to be maintained at 20⁰C and 50⁰C, as taught by Liu, as temperatures in this range allow for separation and resolution of oligonucleotides (Liu, [see Figure 4, page 371]). It has been held that obviousness exists where the claimed ranges overlap or lie inside ranges disclosed by the prior art. See MPEP 2144.05 (I). Furthermore, generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. “[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." MPEP § 2144.05(II)(A).
Regarding Claim 2, modified Thanthri teaches the method of claim 1.
Thanthri teaches wherein the first reservoir solution includes a first electrolyte (trailing electrolyte solution contains glycine [third para. col. 2, page 6742]) and the second reservoir solution includes a second electrolyte (leading electrolyte solution contains tris-HCl [third para. col. 2, page 6742]).
Regarding Claim 3, modified Thanthri teaches the method of claim 1.
Thanthri teaches wherein the microfluidic device further comprises a first electrode arranged in the first reservoir (as illustrated in Figure 1 on page 6742, a first electrode applying 0 kV is arranged in the first reservoir) and a second electrode arranged in the second reservoir (as illustrated in Figure 1 on page 6742, a second electrode applying +2 kV is arranged in the second reservoir).
Regarding Claim 4, modified Thanthri teaches the method of claim 3.
Thanthri teaches wherein: the method comprises anionic analytes migrating from the first reservoir to the second reservoir (ovalbumin migrated along the microchannel towards the LE reservoir (i.e., from the first reservoir to the second reservoir) [second para. col. 2, page 6743]), and:
the first electrode is a cathodic electrode (as illustrated in Figure 1, first electrode is a cathodic electrode with a voltage of 0 kV applied [page 6742]);
the first reservoir solution is a cathodic reservoir solution (trailing electrolyte solution contains 100 mM Glycine and 50 mM Tris-HCl serves as a cathodic reservoir solution [third para. col. 2, page 6742]);
the first reservoir is a cathodic reservoir (as the electrode in the trailing electrolyte is applying no voltage compared to the electrode applying a positive voltage in the leading electrolyte, the leading electrolyte reservoir serves as a cathodic reservoir [see Figure 1 on page 6742]
the second electrode is an anodic electrode (as illustrated in Figure 1, second electrode is an anodic electrode with a voltage of +2 kV applied [page 6742];
the second reservoir solution is an anodic reservoir solution (leading electrolyte solution contains 25 mM Tris-HCl and 30% PF-127 [third para. col. 2, page 6742]); and
the second reservoir is an anodic reservoir (as the electrode in the leading electrode is applying a positive voltage compared to the electrode applying no voltage in the trailing electrolyte, the leading electrolyte reservoir serves as an anodic reservoir [see Figure 1 on page 6742]).
Regarding Claim 6, modified Thanthri teaches the method of claim 1.
Thanthri teaches wherein one or more of:
the mixed analyte sample comprises biomolecules (ovalbumin-Texas Red dye conjugate protein [third para. col. 1, page 6742]);
the mixed analyte sample comprises at least one of proteins (ovalbumin-Texas Red dye conjugate protein [third para. col. 1, page 6742]);
the gel is thermally responsive (Pluronic F-127, which is a thermally reversible polymer [first para. col. 2, page 6741]).
Regarding Claim 15, modified Thanthri teaches the method of claim 1.
Thanthri teaches further comprising: including buffer in the gel solution (sample-containing gels were prepared for analysis by diluting with 5 mM Tris-HCl [third para. col. 2, page 6742]) and/or the mixed analyte sample (protein stock solution was diluted with 1X phosphate-buffered saline [third para. col. 1, page 6742]); and/or detecting separation of analytes of the mixed analyte sample in the channel (separation of different ovalbumin protein variants are seen in images in Figure 2B [page 6743]).
Regarding Claim 21, modified Thanthri teaches the method of claim 2.
Thanthri teaches wherein at least the first electrolyte is glycine or tris-HCl (trailing electrolyte solution contains 100 mM Glycine and 50 mM Tris-HCl [third para. col. 2, page 6742]) or at least the second electrolyte is Tris-HCl (leading electrolyte solution contains 25 mM Tris-HCl [third para. col. 2, page 6742]).
Regarding Claim 22, modified Thanthri teaches the method of claim 4.
Thanthri teaches wherein: the cathodic reservoir solution comprises at least one of glycine or Tris-HCl (trailing electrolyte solution contains 100 mM Glycine and 50 mM Tris-HCl [third para. col. 2, page 6742]; and, as illustrated in Figure 2, the TE reservoir serves as the cathodic reservoir, [page 6742]);
or the anodic reservoir solution comprises at least one of Tris-HCl (leading electrolyte solution contains 25 mM Tris-HCl [third para. col. 2, page 6742]); and, as illustrated in Figure 2, the LE reservoir serves as the anodic reservoir [page 6742]).
Regarding Claim 24, modified Thanthri teaches the method of claim 2.
Thanthri teaches wherein the first reservoir solution includes at least two different electrolyte species (trailing electrolyte solution contains 100 mM Glycine and 50 mM Tris-HCl [third para. col. 2, page 6742]).
Regarding Claim 29, modified Thanthri teaches the method of claim 6.
Thanthri teaches wherein the mixed analyte sample comprises at least one biomolecule that occurs in two or more different conformations (Fluorescently labeled ovalbumin was selected as the analyte for native protein because it has multiple folding conformations [second para. col. 1, page 6743]) each of which has a different electrophoretic mobility (as illustrated in Figure 2B, different conformations of ovalbumin are separated by thermal gel electrophoresis based on electrophoretic mobility [page 6743]); and optionally the method separates two or more different conformational forms of the protein (Figure 2B shows different conformations of ovalbumin separated in the microchannel [page 6743]).
Claim 26 is rejected under 35 U.S.C. 103 as being unpatentable over Thanthri and Liu. Beckman User Manual (Beckman Coulter P/ACE MDQ User’s Guide, 2009) is used as an evidence reference for claim 26.
Regarding Claim 26, modified Thanthri teaches the method of claim 1.
Although Thanthri does not explicitly teach wherein applying the electric field across the microfluidic device comprises applying the electric field across the microfluidic device at a voltage of:
-10kV to +10kV; -8kV to +8kV; -5kV to +5kV; -3 kV to +3kV; -2 kV to +2kV; -1.5kV to +1.5kV; -1.0 kV to +1.0 kV; -0.5 kV to +0.5 kV; -0.25 kV to +0.25 kV; 1 kV to 2 kV; 1.5 kV to 2 kV; 1.5 kV to 2 kV; 0.5 kV to 1.5 kV; 0.5 kV to 2 kV; 0.5 kV to 1 kV; -1 kV to -2 kV; -1.5 kV to -2 kV; -0.5 kV to -1.5 kV; -0.5 kV to -2 kV; or -0.5 kV to -1 kV,
Thanthri does teach applying a 0 kV and +2 kV voltage for a total applied voltage of 2 kV (Figure 1, page 6742) across a 3 cm microchannel (second para. col. 2, page 6742) so that the electric field along the microchannel is 666.67 V/cm-1 (first para. col. 1, page 6743). From the above claimed voltage spreads, a voltage spread of -1.0 kV to +1.0 kV gives the same total applied voltage of 2 kV for the same electric field of 666.67 V/cm-1. Varying the electric field using voltage is well known in the field of electrophoresis as evidenced by Beckman User Manual (Voltage applied to a capillary channel ranges from 1 to 30 kV [Section 2.3 High Voltage Power Supply, page 8]).
It would be obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to apply the electric field across the microfluidic device of modified Thanthri at a voltage of -1 kV to +1 kV, as a voltage spread of -1 kV to +1 kV gives an equivalent electric field strength of 666.67 V/cm-1, which successfully separated conformations of ovalbumin proteins (Thanthri, [Figure 2B on page 6743]).
Claims 17-18 are rejected under 35 U.S.C. 103 as being unpatentable over Thanthri and Liu, as applied to claim 1 above, and in view of Zhu (Simulation and experiment of asymmetric electrode placement for electrophoretic exclusion in a microdevice, Electrophoresis, 2019; 40, pages 304-314).
Regarding Claim 17, modified Thanthri teaches the method of claim 3.
Thanthri is silent on comprising applying the electric field across the microfluidic device as an asymmetric electric field.
Zhu teaches applying electric fields in microfluidic devices (abstract), and teaches comprising applying the electric field across the microfluidic device as an asymmetric electric field (by varying the location of electrodes, such as electrodes placed outside a channel entrance, electrodes aligned with a channel entrance, and electrodes within the channel, an asymmetric electric field can be applied to a microfluidic channel [last para. col. 2, page 305 to first para. col. 1, page 306].
Modified Thanthri and Zhu are considered analogous art to the claimed inventions because they are in the same field of methods of electrophoresis using microfluidic devices. It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the second electrode to be offset from the channel/second reservoir of modified Thanthri so that applying the electric field across the microfluidic device is an asymmetric field, as taught by Zhu, as asymmetric electric fields allow for more efficient microfluidic interfaces as well as enhanced concentration ability and resolving power (Zhu, [abstract, page 304; concluding remarks, page 313]).
Regarding Claim 18, modified Thanthri method of claim 17, and wherein applying the asymmetric electric field across the microfluidic device comprises applying the asymmetric electric field with the second electrode arranged at an offset position relative to the second reservoir (as outlined in the claim 17 rejection above).
Claim 25 is rejected under 35 U.S.C. 103 as being unpatentable over Thanthri and Liu, as applied to claim 24 above, and in view of Somaweera (Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ether Dye on a PDMS Microfluidic Chip, Micromachines, 2020; 40, pages 1-11).
Regarding Claim 25, modified Thanthri teaches the method of claim 24.
Thanthri is silent on wherein the at least two different electrolyte species comprise glycine and tricine, glycine and borate, or glycine and proline.
Somaweera teaches a method of using isotachophoresis on a microfluidic chip (abstract), and teaches wherein the electrolyte species is composed of borate (trailing electrolyte species contains 200 mM borate electrolyte [first para., page 3]). Note that, as outlined in the claim 24 rejection above, Thanthri teaches the trailing electrolyte comprises glycine.
Modified Thanthri and Somaweera are considered analogous art to the claimed inventions because they are in the same field of methods of electrophoresis using microfluidic devices. It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the trailing electrolyte of modified Thanthri by adding borate so that the at least two different electrolyte species comprise glycine and borate, as taught by combined Thanthri and Somaweera, as adjusting the composition of the trailing electrolyte is common in electrophoresis to obtain maximum preconcentration and resolution of analyte bands (Somaweera, [abstract]).
Claim 59 is rejected under 35 U.S.C. 103 as being unpatentable over Thanthri and Liu, as applied to claim 1 above, and in view of Zhang (A multiple-capillary electrophoresis system for small-scale DNA sequencing and analysis. Nucleic Acids Research 1999; 27(24), pages i-vii).
Regarding Claim 59, modified Thanthri teaches the method of claim 1.
Thanthri is silent on wherein the channel has a tapered geometry.
Zhang teaches a capillary system for DNA sequencing (abstract), and teaches wherein the channel has a tapered geometry (chamber is tapered at the bottom near the detection zone [second para. col. 2, page ii; see also Figure 2, page ii]).
Modified Thanthri and Zhang are considered analogous art to the claimed inventions because they are in the same field of methods of electrophoresis using microfluidic devices. It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify the channel of modified Thanthri to have a tapered geometry, as taught by Zhang, as a tapered geometry allows for successful separation of DNA lengths for applications including sequencing (Zhang, [Figure 5]).
Response to Arguments
Applicant's arguments, see Remarks pgs. 9-10, filed 10/21/2025, with respect to the 35 U.S.C 102 and 35 U.S.C 103 rejections and amended claims have been fully considered.
Applicant’s Argument #1:
Applicant argues and traverses on page 9 that the 35 U.S.C. 102 rejection, as claim 1 has been amended to specify the temperature at which the device is maintained, which is not taught by Thanthri. In addition, Applicant asserts that Thanthri teaches away as Thanthri states “[w]e did not want to risk altering protein folding by heating about physiological temperature (37⁰C)…”.
Examiner’s Response #1:
Applicant’s arguments have been fully considered, but are moot in view of the new grounds of rejection.
Applicant’s Argument #2:
Applicant argues on pages 9-10 that the U.S.C. 103 rejections for dependent claims 26, 17-18, and 25 are traversed as the secondary references of Beckman User Manual, Zhu, and Somaweera do not cure the deficiencies of amended claim 1.
Examiner’s Response #2:
Applicant’s arguments have been fully considered, but are moot in view of the new grounds of rejection.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/R.L.G./Examiner, Art Unit 1795
/LUAN V VAN/Supervisory Patent Examiner, Art Unit 1795