Claims for Patent: 9,868,973
✉ Email this page to a colleague
Summary for Patent: 9,868,973
Title: | Method for optimizing post-translational modifications on recombinant proteins |
Abstract: | A method for optimizing post-translational modifications of recombinant proteins expressed in living cells is described. More particularly, a method for modulation of host proteins in living cells that control PTMs on recombinant proteins is described that has particularly useful applications in developing manufacturing process changes or in biosimilar development. The goal of this modulation is to produce a recipe for production of a recombinant protein in the new process or in the biosimilar that will produce a targeted PTM profile in the resulting protein product. In the method one or more modulators are selected, as from a modulator library, which affect the activity of host proteins. These modulators are added to media during production such that the resulting product matches the PTMs of the reference product. The ideal set of modulators and their concentrations are identified through a unique iterative process and the combined modulators and their concentrations constitute a recipe for growth media for the production of said recombinant protein. The methodology to obtain such a recipe described herein may then be used in many applications, such as optimizing new batches of recombinant protein drugs, developing biosimilar or bio-better drugs. |
Inventor(s): | Leszcyniecka; Magdalena (Cambridge, MA), Shulga-Morskoy; Sergey (Cambridge, MA) |
Assignee: | STC Biologics, Inc. (Cambridge, MA) |
Application Number: | 15/111,260 |
Patent Claims: | 1. A method for producing a biosimilar protein by optimizing a growth medium for living cells that produce a recombinant protein having a target set of post-translational
modifications, said method comprising: a) measuring post-translational modifications on a reference product protein, b) setting target post-translational modification ranges for the recombinant protein based on the measured post-translational
modifications of the reference product protein, c) providing a living host cell that expresses the recombinant protein, d) placing said host cells in growth media containing a first set of post-translational modulators having a first set of
concentrations, e) identifying the post-translational modifications on the recombinant protein and determining the differences between the identified post-translational modifications and the target post-translational modification, ranges based on the
reference product protein as determined in step b), and if the target post-translational modification of the recombinant protein are not within the target post-translational modification ranges determined in b), then: f) developing a second set of
post-translational modulators wherein the second set of post-translational modulators may include a different set of modulators from the first set, different concentrations of modulators from the first set, or a combination of different modulators and
different concentrations of modulators, g) producing the recombinant protein from the cells engineered to express the recombinant protein in a growth media in the presence of the modified second set of post-translational modulators, h) isolating the
recombinant protein from growth media and comparing its post-translational modifications to the target set of post-translational modifications, i) repeating steps e), f), g), and h) until a set of post-translational modulators is identified that can be
used to produce a recombinant protein having the target set of post-translational modulations.
2. The method of claim 1, wherein the post-translational modifications on the recombinant protein are the same as the modifications on the reference protein. 3. The method of claim 1, wherein the post-translational modifications on the recombinant protein match the target post-translational modification. 4. A method of making a trastuzumab biosimilar with same or similar post-translational modifications to a reference trastuzumab product said method comprising: a) measuring post-translational modifications on different batches of trastuzumab reference product, b) setting target post-translational modification ranges for a trastuzumab biosimilar based on the measured post-translational modifications in the reference trastuzumab reference product, c) providing a living host cell engineered to express the trastuzumab, d) placing said host cell in a cell culture medium in the presence of a first set of post-translational modulators having a first set of concentrations, e) identifying the post-translational modifications on the trastuzumab biosimilar protein and determining the differences between the identified post-translational modifications and the target post-translational modification, ranges based on the reference product protein as determined in step b), f) developing a modified second set of post-translational modulators wherein the modified second set of post-translational modulators may include a different set of modulators from the first set, different concentrations of modulators from the first set, or a combination of different modulators and different concentrations of modulators, g) producing the trastuzumab biosimilar protein from the cells engineered to express trastuzumab in a cell culture medium containing the modified second set of post-translational modulators, h) isolating the trastuzumab protein grown in the presence of the modified second set of post-translational modulators from the cell culture and comparing its post-translational modifications to the target set of post-translational modifications, i) repeating steps e), f), g), and h) until a set of post-translational modulators is identified that can be used to produce trastuzumab having the target set of post-translational modifications. 5. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab having the target set of post-translational modifications comprises fucosyltransferase inhibitor. 6. The method of claim 5, wherein the post-translational modulators identified for producing a trastuzumab having the target set of post-translational modifications consists of a fucosyltransferase inhibitor. 7. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications comprises a fucosyltransferase and an a-mannosidase I inhibitor. 8. The method of claim 7, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications consists of a fucosyltransferase and an a-mannosidase I inhibitor. 9. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications comprises 2F-Peracetyl-Fucose. 10. The method of claim 9, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications consists of 2F-Peracetyl-Fucose. 11. The method of claim 9, wherein the 2F-Peracetyl-Fucose in the growth media is about 0.1 M to about 20 M. 12. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications comprises a GDP-Fucose derivative. 13. The method of claim 12, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target post-translational modifications consists of a GDP-Fucose derivative. 14. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target post-translational modifications comprises a specific inhibitor of .alpha.-mannosidase I. 15. The method of claim 4, wherein the post-translational modulators identified for producing a trastuzumab biosimilar having the target set of post-translational modifications comprises kifunensine. 16. The method of claim 15, wherein the kifunensine in the growth media is about 0.01ng/ml to about 20 ng/ml. 17. The method of claim 4, wherein the mammalian cell is a hybridoma, a CHO cell, CAP-T cells, 293 or an NSO cell. |
Details for Patent 9,868,973
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Genentech, Inc. | HERCEPTIN | trastuzumab | For Injection | 103792 | 09/25/1998 | ⤷ Try a Trial | 2039-02-26 |
Genentech, Inc. | HERCEPTIN | trastuzumab | For Injection | 103792 | 02/10/2017 | ⤷ Try a Trial | 2039-02-26 |
Genentech, Inc. | HERCEPTIN HYLECTA | trastuzumab and hyaluronidase-oysk | Injection | 761106 | 02/28/2019 | ⤷ Try a Trial | 2039-02-26 |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
Make Better Decisions: Try a trial or see plans & pricing
Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.