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1. (WO2019050856) METHODS AND COMPOSITIONS FOR PREPARING SURFACTANT PROTEIN D (SP-D)
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WHAT IS CLAIMED IS:

1. A composition comprising a human surfactant protein D (SP-D) polypeptide prepared by a method comprising:

(a) introducing a polynucleotide encoding the SP-D polypeptide into a human mammalian cell;

(b) culturing the cell under conditions in which the SP-D polypeptide is expressed; and

(c) isolating the expressed SP-D polypeptide from the cell.

2. The composition of claim 1, wherein the cell is derived from a human myeloid leukemia cell.

3. The composition of claim 2, wherein the cell is selected from the group consisting of M-H9D8, M-H9D8-E6Q12, and M-F9.

4. The composition of claim 3, wherein the cell is a M-H9D8 cell.

5. The composition of claim 3, wherein the cell is a M-H9D8(8B11) cell.

6. The composition of claim 1, wherein the polynucleotide encodes a wild type SP-D polypeptide leader sequence.

7. The composition of claim 6, wherein the polynucleotide comprises SEQ ID

NO:03.

8. The composition of claim 6, wherein the polynucleotide comprises SEQ ID

NO:02.

9. The composition of claim 6, wherein the polynucleotide encodes a polypeptide having an amino acid sequencing comprising SEQ ID NO:05.

10. The composition of claim 6, wherein the polynucleotide encodes a polypeptide having an amino acid sequencing comprising SEQ ID NO:04.

11. The composition of claim 1, wherein the polynucleotide encodes a wild type T-cell receptor (TCR) polypeptide leader sequence.

12. The composition of claim 11, wherein the polynucleotide comprises SEQ ID

NO:08.

13. The composition of claim 11, wherein the polynucleotide comprises SEQ ID

NO:07.

14. The composition of claim 11, wherein the polynucleotide encodes a polypeptide having an amino acid sequencing comprising SEQ ID NO: 10.

15. The composition of claim 11, wherein the polynucleotide encodes a polypeptide having an amino acid sequencing comprising SEQ ID NO:09.

16. The composition of claim 1, wherein the SP-D polypeptide comprises residue at a polymorphic position, wherein the residue is selected from the group consisting of Metl 1/31, Thrl60/180, Ser 270/290, and Ala 286/306.

17. The composition of claim 16, wherein the SP-D polypeptide comprises Metl 1/31.

18. The composition of claim 16, wherein the SP-D polypeptide comprises Metl 1/31, Thrl60/180, Ser 270/290, and Ala 286/306.

19. The composition of claim 1, wherein the human SP-D polypeptide comprises the amino acid sequence of positions 22 to 376 of SEQ ID NO:04.

20. The composition of claim 1 comprising a population of the expressed SP-D polypeptides, each expressed SP-D polypeptide comprising a complex-type carbohydrate attached at an N-glycosylation site, wherein the population has a glycosylation pattern comprising the following characteristics:

(i) at least 70% of the complex-type carbohydrates include a core fucose;

(ii) at least 10% of the complex-type carbohydrates include at least one sialic acid residue;

(iii) at least 50% of the complex-type carbohydrates include at least a biantennary carbohydrate structure;

(iv) at least 10% of the complex-type carbohydrates include a bisecting N- acetylglucosamine;

(v) less than 10% of the carbohydrates are high-mannose type structures; and

(vi) a detectable amount of a2,6-coupled sialic acid residues.

21. The composition of claim 20, wherein the population has a glycosylation pattern comprising one or more of the following characteristics:

(i) at least 20% of the complex-type carbohydrates include a bisecting N- acetylglucosamine; and

(ii) at least 85% of the complex-type carbohydrates include a core fucose.

22. The composition of claim 1, wherein the polynucleotide encodes a dihydrofolate reductase polypeptide.

23. The composition of claim 1, wherein culturing the cell comprises contacting the cell with an antifolate.

24. The composition of claim 23, wherein expression of the SP-D polypeptide is increased by increasing the concentration of the antifolate.

25. The composition of claim 23, wherein the antifolate comprises methotrexate.

26. The composition of claim 1, wherein the cell is cultured in a perfusion bioreactor.

27. The composition of claim 1, wherein the cell is cultured in a continuous culture.

28. The composition of claim 1, wherein culturing the cell comprises maintaining a growth medium having a pH 7.2, dissolved oxygen at 40% and or 20%, and temperature at 37°C.

29. The composition of claim 28, wherein the dissolved oxygen is lower than 35%, preferably 30%.

30. The composition of claim 1, wherein isolating the expressed SP-D polypeptide from the cell comprises preparing a cell supernatant from a culture medium containing the cell.

31. A solution comprising a population of recombinant human surfactant protein D (SP-D) polypeptides having oligomeric forms, wherein greater than about 40% of the oligomeric forms comprise dodecamers of the SP-D.

32. The solution of claim 31, wherein the human SP-D polypeptide comprises the amino acid sequence of positions 22 to 376 of SEQ ID NO:04.

33. The solution of claim 31, wherein the SP-D polypeptide comprises residue at a polymorphic position, wherein the residue is selected from the group consisting of Metl 1/31, Thrl60/180, Ser 270/290, and Ala 286/306.

34. The solution of claim 31, wherein the SP-D polypeptide comprises Metl 1/31.

35. The solution of claim 31, wherein the SP-D polypeptide comprises Metl 1/31, Thrl60/180, Ser 270/290, and Ala 286/306.

36. The solution of claim 31, wherein greater than about 60% of the oligomeric forms comprise dodecamers of the SP-D.

37. The solution of claim 31, wherein greater than about 62% of the oligomeric forms comprise dodecamers of the SP-D.

38. The solution of claim 31, wherein greater than about 64% of the oligomeric forms comprise dodecamers of the SP-D.

39. The solution of claim 31, wherein a distribution of oligomeric forms of the SP-D is measured using an asymmetric flow field-flow fractionation with multi-angle light scattering (AF4-MALS) analysis.

40. The solution of claim 31, wherein each SP-D polypeptide comprises a complex-type carbohydrate attached at an N-glycosylation site, wherein the population has a glycosylation pattern comprising the following characteristics:

(i) at least 70% of the complex-type carbohydrates include a core fucose;

(ii) at least 10% of the complex-type carbohydrates include at least one sialic acid residue;

(iii) at least 50% of the complex-type carbohydrates include at least a biantennary carbohydrate structure;

(iv) at least 10% of the complex-type carbohydrates include a bisecting N- acetylglucosamine;

(v) less than 10% of the carbohydrates are high-mannose type structures; and

(vi) a detectable amount of a2,6-coupled sialic acid residues.

41. The solution of claim 40, wherein the population has a glycosylation pattern comprising one or more of the following characteristics:

(i) at least 20% of the complex-type carbohydrates include a bisecting N- acetylglucosamine; and

(ii) at least 85% of the complex-type carbohydrates include a core fucose.

42. A composition comprising a human surfactant protein D (SP-D) polypeptide prepared by a method comprising:

(a) culturing a human mammalian comprising an introduced cell polynucleotide encoding the SP-D polypeptide under conditions in which the SP-D polypeptide is expressed; and

(b) isolating the expressed SP-D polypeptide from the cell.

43. The composition of claim 42, wherein the cell is derived from a human myeloid leukemia cell.

44. The composition of claim 43, wherein the cell is selected from the group consisting of M-H9D8, M-H9D8-E6Q12, and M-F9.

45. The composition of claim 44, wherein the cell is a M-H9D8 cell.

46. The composition of claim 42, wherein the cell is a M-H9D8(8B11) cell.