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 .
Acknowledgement of Receipt
Applicant’s Response, filed 6/18/2026, in reply to the Office Action mailed 3/20/2026, is acknowledged and has been entered. Claims 1-3, 5-12, 14 and 15 are pending, of which claims 6-8, 11, 12, 14 and 15 are withdrawn from consideration at this time as being drawn to a non-elected invention. Claims 1-3, 5, 9 and 10 encompass the elected invention and are examined herein on the merits for patentability.
Response to Arguments
Applicant’s arguments have been fully considered. The rejections over Gatzinksy et al. (Brain Research, 2001, 920: 226-238) and Kasaian et al. (Exp. Cell. Res., 1994, 210: 77-85) have been withdrawn in view of Applicant’s showing that Gatzinsky and Kasaian’s proteins are in dimeric form. Applicant's request for reconsideration of the finality of the rejection of the last Office action is persuasive and, therefore, the finality of that action is withdrawn. New grounds of rejection are set forth herein, made final in view of the amendment filed 12/18/2025.
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claim(s) 1-3, 5, 9 and 10 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by De Nadai et al. (Scientific Reports, 2016, 6, 20272).
De Nadai discloses that the classical view of action in the nervous system is linked to its retrograde nerve growth factor (NGF) axonal transport. However, almost nothing is known on the trafficking properties of its unprocessed precursor proNGF, characterized by different and generally opposite biological functions with respect to its mature counterpart. Here we developed a strategy to fluorolabel both purified precursor and mature neurotrophins (NTs) with a controlled stoichiometry and insertion site. Using a single particle tracking approach, we characterized the axonal transport of proNGF versus mature NGF in living dorsal root ganglion neurons grown in compartmentalized microfluidic devices. We demonstrate that proNGF is retrogradely transported as NGF, but with a lower flux and a different distribution of numbers of neurotrophins per vesicle. Moreover, exploiting a dual-color labelling technique, we analysed the transport of both NT forms when simultaneously administered to the axon tips.
A novel fluorolabeling strategy is taught, allowing for the production of “homologous” fluorescent human NGF and proNGF, based on the insertion of an 11 amino acid tag at the at the C- terminus of the protomer sequence. The inserted tag is a target for a site-specific enzymatic covalent binding and it is used here to bind a small organic dye, so that a site-specific fluorophore conjugation with 1:1 (label:NT-monomer) stoichiometry is obtained for both the unprocessed and mature forms of NGF. The technique allows for a high (≈80%) fluorescent NT production yield and for an optimal purification from the unlabeled counterparts. The obtained labelled species retain the same functional features of the native proteins. Fluorescence microscopy experiments were performed on compartmentalized living cultures of rat dorsal root ganglion (DRG) neurons, in which fluorescent proNGF and NGF were administered either separately or together. The first direct evidence that proNGF is retrogradely transported like mature NGF is provided here, although important differences of the axonal transport of the two molecules have been uncovered. Crucially, the controlled stoichiometry of the labelling reaction allowed quantifying the number of NTs in each vesicle, showing a significant difference between their distributions in the two cases. Moreover, by coadministering both neurotrophins labelled with different fluorophores, we were able to analyse the cotransport of precursor and mature neurotrophins in neurons (page 1-2).
10 µg of proNGF-YBBR or NGF-YBBR were incubated for 30 minutes at 37 °C in a thermomixer at 300 rpm with a reaction mix (10 mM MgCl2, 10 µM CoA-alexa488/ CoA-alexa 647 or CoA-biotin and 2 µM Sfp Synthase (SfpS) (New England Biolabs), in phosphate buffer up to 250 µl final volume (Experimental).
In Figure 1, the tag sequence inserted at the C-terminal position of proNGF is depicted in red (Middle); the complete structural formula of Alexa488-maleimide-phosphopantetheinyl is added, highlighted in green.
Conclusion
No claims are allowed at this time.
THIS ACTION IS MADE FINAL. 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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to LEAH H SCHLIENTZ whose telephone number is (571)272-9928. The examiner can normally be reached Monday-Friday, 8:30am - 12:30pm EST.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, MICHAEL HARTLEY can be reached at 571-272-0616. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/LHS/
/Michael G. Hartley/Supervisory Patent Examiner, Art Unit 1618
Prosecution Insights