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Table 3 Purification of insulin precursor by IMAC and conversion to human insulin

From: Application of simple fed-batch technique to high-level secretory production of insulin precursor using Pichia pastoris with subsequent purification and conversion to human insulin

Steps

Insulin species

(molecular mass)

Amount (mg)a

Amount (mmol)a

Recovery (%)a

Purity (%)a

Culture broth b

Insulin precursor

7041 Da

3075

0.437

100

20

Chelating sepharose FF c

Insulin precursor

7041 Da

2921

0.415

95

96

Desalting & Lyophilization

Insulin precursor

7041 Da

2859

0.406

93

98

Transpeptidation

Pre-insulin

5920 Da

n.d. d

n.d. d

n.d. d

n.d. d

Resource RPC Desalting & Lyophilization

Pre-insulin

5920 Da

1915

0.323

74

90

Deprotection

Human insulin

5808 Da

n.d. d

n.d. d

n.d. d

n.d. d

Resource RPC (two passages), Desalting & Lyophilization

Human insulin

5808 Da

1537

0.264

60

99

  1. a Recovery and purity was determined by RP-HPLC
  2. b 200 mg of IP were used for further processing. Data extrapolated to 1-L culture broth
  3. c Centrifugation step included before chromatography
  4. d n.d., not determined