Antibodies With Modulated Glycan Profiles
Patent No. US12084686 (titled "Antibodies With Modulated Glycan Profiles") was filed by Amgen Inc on Apr 30, 2019.
What is this patent about?
’686 is related to the field of recombinant antibody production, specifically addressing the modulation of glycan profiles on therapeutic antibodies like denosumab. The background acknowledges that the glycosylation patterns of antibodies significantly impact their safety, efficacy, immunogenicity, and half-life. Prior art struggles to control the levels of high-mannose glycans, which can affect therapeutic activity and clearance rates, creating a need for methods to fine-tune these glycan profiles to achieve a desired balance between pharmacokinetic properties and therapeutic effects.
The underlying idea behind ’686 is to control the high-mannose glycan content of recombinantly produced denosumab by manipulating the glucose and galactose concentrations in the cell culture medium. The key inventive insight is that by reducing glucose and increasing galactose during the production phase, the level of high-mannose glycans, particularly Mannose-5 (Man-5), attached to the denosumab molecule at the N-298 site can be increased. This is achieved without significantly compromising cell growth, viability, or overall antibody production.
The claims of ’686 focus on a method for increasing the level of high-mannose present on denosumab molecules, where the denosumab molecules are recombinantly expressed by a mammalian host cell. The method involves two key steps: (a) incubating the host cell in a first culture medium during the growth phase until a certain cell density is reached, with a specified glucose concentration range; and (b) subsequently incubating the host cells in a second culture medium during the production phase, with a lower glucose concentration range and a specified galactose concentration range. The independent claim specifies that the resulting denosumab molecules will have a high-mannose glycan content at the N-298 site within a range of 2% to 14%.
In practice, the method involves culturing mammalian cells, such as CHO cells, in a bioreactor. The cells are first grown in a glucose-rich medium to achieve a high cell density. Then, the culture medium is switched to a medium with reduced glucose and supplemented with galactose. This change in the carbon source forces the cells to utilize galactose for glycosylation, leading to an increase in the proportion of high-mannose glycans on the produced denosumab. The glucose and galactose concentrations are carefully controlled, often through bolus feeds or perfusion, to maintain optimal cell health and antibody production.
This approach differentiates from prior art by directly addressing the need for controlled modulation of high-mannose content. Unlike previous methods that might rely on enzymatic treatments or genetic modifications, ’686 provides a relatively simple and controllable method based on adjusting the culture medium composition. This allows for a more precise tuning of the glycan profile, potentially leading to improved therapeutic efficacy or pharmacokinetic properties of the denosumab product. The use of galactose as an alternative carbon source is a key aspect of this differentiation, enabling the targeted increase in high-mannose glycans.
How does this patent fit in bigger picture?
Technical landscape at the time
In the late 2010s when ’686 was filed, at a time when therapeutic antibodies were typically produced using mammalian cell culture, controlling glycosylation patterns was a known challenge, especially when hardware or software constraints made precise control of culture conditions non-trivial.
Novelty and Inventive Step
The examiner approved the claims because they are directed to a method of increasing the level of high-mannose present on denosumab molecules, wherein said denosumab molecules are recombinantly-expressed by a mammalian host cell, comprising: (a) incubating said mammalian host cell in a first culture medium during growth phase until the cell density is at least 1x106 viable cells/mL, wherein said first culture medium comprises from 1 g/L to 20 g/L glucose; and subsequently (b) incubating host cells from step (a) in a second culture medium during production phase to express said denosumab molecules, wherein said second culture medium comprises from 0 g/L to 10 g/L glucose and from 5 g/L to 20 g/L galactose; wherein from 2% to 14% of the denosumab molecules comprise high-mannose glycan at N-298 site. The examiner stated that the claimed method is free of prior art and that the prior art fails to suggest such specifically claimed method. Hence, said method is also non-obvious.
Claims
This patent contains 11 claims, with claims 1 and 1 being independent. The independent claims are directed to methods of increasing the level of high-mannose present on denosumab molecules recombinantly-expressed by a mammalian host cell. The dependent claims generally narrow the scope of the independent claims by specifying details such as glucose and galactose concentrations, cell densities, culture media, and cell types.
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Date
Description
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US12084686
- Application Number
- US17051528
- Filing Date
- Apr 30, 2019
- Status
- Granted
- Expiry Date
- Sep 23, 2041
- External Links
- Slate, USPTO, Google Patents