Human growth hormone and insulin, from tissue extracts to recombinant DNA technology, their biosimilars and analogues in treatment of Endocrine Disorders

Main Article Content

Linda Fryklund, PhD


Forty years ago, recombinant DNA technology caused a revolution in the way growth disorders and diabetes could be treated. Bacteria or human cells were modified so as to be able to produce human hormones instead of relying on cadaveric material in the case of growth disorders and porcine insulin in the case of diabetics. The new technology also removed the risk of contracted hepatitis or fatal Creutzfeldt Jacob disease.

A signal peptide was used to transport the new protein into the periplasmic space giving rise to an exact, correctly folded version of monomeric human growth hormone.

More eligible patients could be treated, resulting not only in a large number of clinical trials. Daily tiny subcutaneous injections were found to be much more effective than twice or thrice weekly in the treatment of children and adults,

The progress for recombinant human insulin turned out to be somewhat different. Insulin is a more complicated protein, with an Alpha and Beta chain joined by two disulphide bridges, but is produced in the human pancreas as a single chain, proinsulin, the C peptide is removed to make the active molecule.

Recombinant insulin development was in a way the opposite to growth hormone since human insulin acts faster than porcine. Short fast and long-acting analogues have been developed all to improve the treatment regimens for both type 1 and type 2 diabetics,

A similar development is now taking place in the growth hormone field. Depot preparations for weekly administration are under development to achieve better compliance. One recently approved depot uses an analogue, modified in one amino acid of the peptide chain to permit acylation and albumin binding. In a way this is a retrograde step and it remains to be seen if the patient’s growth and metabolic responses are comparable to the daily administration of tiny amount of recombinant natural sequence hormone over several years of treatment.

Article Details

How to Cite
FRYKLUND, Linda. Human growth hormone and insulin, from tissue extracts to recombinant DNA technology, their biosimilars and analogues in treatment of Endocrine Disorders. Medical Research Archives, [S.l.], v. 11, n. 3, mar. 2023. ISSN 2375-1924. Available at: <>. Date accessed: 20 apr. 2024. doi:
Research Articles


1. Fryklund L. Production of authentic recombinant somatropin. Acta Paediatr Scand Suppl. 1987;337:134. doi: 10.1111/j.1651-2227.1987.tb17147.x.PMID: 3481180 No abstract available.
2. Gellerfors P, Eketorp G, Fhölenhag K, Pavlu B, Johansson S, Fryklund L. Characterisation of a secreted form of recombinant derived human growth hormone, expressed in Escherichia coli cells. J Pharm Biomed Anal. 1989;7(2):173-83. doi: 10.1016/0731-7085(89)80081-0. PMID: 2488618.
3. Riggs AD. Making, Cloning, and the Expression of Human Insulin Genes in Bacteria: The Path to Humulin. Endocr Rev. 2021 May 25;42(3):374-380. doi: 10.1210/endrev/bnaa029.
4. Farrugia A. Plasma for fractionation: safety and quality issues. Haemophilia. 2004 Jul;10(4):334-40. doi: 10.1111/j.1365-2516.2004.00911.x. PMID: 15230946.
5. Isfordink CJ, van Erpecum KJ, van der Valk M, Mauser-Bunschoten EP, Makris M. Viral hepatitis in haemophilia: historical perspective and current management. Br J Haematol. 2021 Oct;195(2):174-185. doi: 10.1111/bjh.17438. Epub 2021 May 6.PMID: 33955555.
6. Collett-Solberg PF, Ambler G, Backeljauw PF, Bidlingmaier M, Biller BMK, Boguszewski MCS, Cheung PT, Choong CSY, Cohen LE, Cohen P, Dauber A, Deal CL, Gong C, Hasegawa Y, Hoffman AR, Hofman PL, Horikawa R, Jorge AAL, Juul A, Kamenický P, Khadilkar V, Kopchick JJ, Kriström B, Lopes MLA, Luo X, Miller BS, Misra M, Netchine I, Radovick S, Ranke MB, Rogol AD, Rosenfeld RG, Saenger P, Wit JM, Woelfle J. Diagnosis, Genetics, and Therapy of Short Stature in Children: A Growth Hormone Research Society International Perspective. Horm Res Paediatr. 2019;92(1):1-14. doi: 10.1159/000502231. Epub 2019 Sep 12. PMID: 31514194; PMCID: PMC6979443.
7. Roos P, Fevold Hr, Gemzell Ca. Preparation Of Human Growth Hormone By Gel Filtration. Biochim Biophys Acta. 1963 Aug 13;74:525-31. doi: 10.1016/0006-3002(63)91395-7. PMID: 14071596
8. Holmström B, Fhölenhag K. Characterization of human growth hormone preparations used for the treatment of pituitary dwarfism: a comparison of concurrently used batches. J Clin Endocrinol Metab. 1975 May;40(5):856-62. doi: 10.1210/jcem-40-5-856. PMID: 1127091
9. Billette de Villemeur T and Pradel A. Iatrogenic Creutzfeldt-Jakob disease. Lessons from cases secondary to extracted growth hormone in France, Transfus Clin Biol. 1994;1(5):333-7.
10. Rudge P 1, Jaunmuktane Z, Adlard P, Bjurstrom N, Caine D, Lowe J, NorsworthyP, Hummerich P Druyeh R, Wadsworth JDF, Brandner S, Hyare H, Mead S, Collinge J. Brain 2015 Nov;138(Pt 11):3386-99.doi: 10.1093/brain/awv235. Epub 2015 Aug 11.
11. Brostedt P, Roos P. Isolation of dimeric forms of human pituitary growth hormone. Prep Biochem. 1989;19(3):217-29. doi: 10.1080/10826068908544912. PMID: 2616540.
12. Purrello F, Vigneri R, Belfiore A, Pezzino V, Squatrito S, Polosa P. High incidence of anti-GH antibodies in subjects treated with the GH clinical preparation available in Italy. J Endocrinol Invest. 1980 Jul-Sep;3(3):313-5. doi: 10.1007/BF03348283. PMID: 7430559.
13. Li, C.H., Human Pituitary Growth Hormone .19. Primary Structure Of Hormone, Archives Of Biochemistry And Biophysics 133: 70 (1969).
14. de Vos AM, Ultsch M, Kossiakoff AA. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science. 1992 Jan 17;255(5042):306-12. doi: 10.1126/science.1549776. PMID: 1549776.
15. Ricci MS, Brems DN. Common structural stability properties of 4-helical bundle cytokines: possible physiological and pharmaceutical consequences. Curr Pharm Des. 2004;10(31):3901-11. doi: 10.2174/1381612043382611. PMID: 15579079.
16. Somers W, Ultsch M, De Vos AM, Kossiakoff AA. The X-ray structure of a growth hormone-prolactin receptor complex. Nature. 1994 Dec 1;372(6505):478-81. doi: 10.1038/372478a0. PMID: 7984244.
17. Baumann G. Growth hormone binding proteins: biochemical characterization and assays. Acta Endocrinol (Copenh). 1991;124 Suppl 2:21-6. PMID: 185367
18. Rubenstein AH, Steiner DF, Cho S, Lawrence AM, Kirsteins L. Immunological properties of bovine proinsulin and related fractions. Diabetes. 1969 Sep;18(9):598-605. doi: 10.2337/diab.18.9.598. PMID: 5821058.
19. Markussen J, Heding LG, Jorgensen KH, Sundby F. Proinsulin, insulin and C-peptide. Horm Metab Res. 1971;3:Suppl 3:33-5. PMID: 5170499.
20. Rançon F, Rosselin G. Etude comparative de l'immuno-réactivité de l'insuline, du C peptide et de la proinsuline de porc avec différents immunsérums anti-proinsuline de porc [Comparative study of immuno-reactivity of swine insulin, C peptide and proinsulin with various anti-proinsulin immune serums of swine]. C R Acad Hebd Seances Acad Sci D. 1972 Apr 5;274(14):2112-5. French. PMID: 4114023.
21. Virgili F, Frigato, F, Magnanini PG, Coronel GA, Iavicoli M. Reduction in insulin antibodies 42 months after transfer from porcine to human monocomponent insulins Diabetes Res 1988 Sep;9(1):19-20.
22. G Schernthaner Affinity of IgG-insulin antibodies to human (recombinant DNA) insulin and porcine insulin in insulin-treated diabetic individuals with and without insulin resistance Diabetes Care. 1982 Nov-Dec;5 Suppl 2:114-8. doi: 10.2337/diacare.5.2.s114
23. Mukherjee S, Sayantan Mondal S, Anilrao Deshmukh A, Gopal B, Bagchi B. What Gives an Insulin Hexamer Its Unique Shape and Stability? Role of Ten Confined Water Molecules Phys Chem B 2018 Feb 8;122(5):1631-1637. doi: 10.1021/acs.jpcb.8b00453. Epub 2018 Jan 27.
24. Gellerfors P, Pavlu B, Axelsson K, Nyhlén C, Johansson S. Separation and identification of growth hormone variants with high performance liquid chromatography techniques. Acta Paediatr Scand Suppl. 1990;370:93-100. doi: 10.1111/j.1651-2227.1990.tb11682.x. PMID: 2260463.
25. Karlsson G, Gellerfors P, Persson A, Norén B, Edlund PO, Sandberg C, Birnbaum S. Separation of oxidized and deamidated human growth hormone variants by isocratic reversed-phase high-performance liquid chromatography. J Chromatogr A. 1999 Sep 3;855(1):147-55. doi: 10.1016/s0021-9673(99)00669-x. PMID: 10514980.
26. Andersson C, Edlund PO, Gellerfors P, Hansson Y, Holmberg E, Hult C, Johansson S, Kördel J, Lundin R, Mendel-Hartvig IB, Norén B, Wehler T, Widmalm G, Ohman J. Isolation and characterization of a trisulfide variant of recombinant human growth hormone formed during expression in Escherichia coli. Int J Pept Protein Res. 1996 Apr;47(4):311-21. doi: 10.1111/j.1399-3011.1996.tb01360.x. PMID: 8738657..
27. Boguszewski MCS. Growth hormone deficiency and replacement in children. Rev Endocr Metab Disord. 2021 Mar;22(1):101-108. doi: 10.1007/s11154-020-09604-2. Epub 2020 Oct 8. PMID: 33029711.Jul 25. PMID: 34304361; PMCID: PMC8416866.
28. Ranke MB, Wit JM. Growth hormone - past, present and future. Nat Rev Endocrinol. 2018 May;14(5):285-300. doi: 10.1038/nrendo.2018.22. Epub 2018 Mar 16. PMID: 29546874.
29. Deodati A, Cianfarani S. The Rationale for Growth Hormone Therapy in Children with Short Stature. J Clin Res Pediatr Endocrinol. 2017 Dec 30;9(Suppl 2):23-32. doi: 10.4274/jcrpe.2017.S003. Epub 2017 Dec 27. PMID: 29280742; PMCID: PMC5790327
30. Richmond E, Rogol AD. Treatment of growth hormone deficiency in children, adolescents and at the transitional age. Best Pract Res Clin Endocrinol Metab. 2016 Dec;30(6):749-755. doi: 10.1016/j.beem.2016.11.005. Epub 2016 Nov 4. PMID: 27974188..
31. Boguszewski MCS, Cardoso-Demartini AA, Boguszewski CL, Chemaitilly W, Higham CE, Johannsson G, Yuen KCJ. Safety of growth hormone (GH) treatment in GH deficient children and adults treated for cancer and non-malignant intracranial tumors-a review of research and clinical practice. Pituitary. 2021 Oct;24(5):810-827. doi: 10.1007/s11102-021-01173-0. Epub 2021 Jul 25. PMID: 34304361; PMCID: PMC8416866
32. de Zegher F, Albertsson-Wikland K, Wilton P, Chatelain P, Jonsson B, Löfström A, Butenandt O, Chaussain JL. Growth hormone treatment of short children born small for gestational age: metanalysis of four independent, randomized, controlled, multicentre studies. Acta Paediatr Suppl. 1996 Oct;417:27-31. doi: 10.1111/j.1651-2227.1996.tb14289.x. PMID: 9055905.
33. Thaker V, Haagensen AL, Carter B, Fedorowicz Z, Houston BW. Recombinant growth hormone therapy for cystic fibrosis in children and young adults. Cochrane Database Syst Rev. 2013 Jun 5;6(6):CD008901. doi: 10.1002/14651858.CD008901.pub2. Update in: Cochrane Database Syst Rev. 2015;(5):CD008901. PMID: 23737090; PMCID: PMC4465600.
34. Dutta D, Mahajan K, Kumar M, Sharma M. Efficacy and safety of long-acting growth hormone in adult growth hormone deficiency: A systematic review and meta-analysis. Diabetes Metab Syndr. 2022 Feb;16(2):102421. doi: 10.1016/j.dsx.2022.102421. Epub 2022 Feb 8. PMID: 35158212

35. Ranke MB, Saenger P. Turner's syndrome. Lancet. 2001 Jul 28;358(9278):309-14. doi: 10.1016/S0140-6736(01)05487-3. PMID: 11498234
36. Steiner M, Saenger P. Turner Syndrome: An Update. Adv Pediatr. 2022 Aug;69(1):177-202. doi: 10.1016/j.yapd.2022.03.004. Epub 2022 Jun 17. PMID: 35985709.
37. Rosenberg AGW, Passone CGB, Pellikaan K, Damiani D, van der Lely AJ, Polak M, Bernardo WM, de Graaff LCG. Growth Hormone Treatment for Adults With Prader-Willi Syndrome: A Meta-Analysis. J Clin Endocrinol Metab. 2021 Sep 27;106(10):3068-3091. doi: 10.1210/clinem/dgab406. PMID: 34105729; PMCID: PMC8475230.
38. Atkinson HF, Moyer RF, Yacoub D, Coughlin D, Birmingham TB. Effects of Recombinant Human Growth Hormone for Osteoporosis: Systematic Review and Meta-Analysis. Can J Aging. 2017 Mar;36(1):41-54. doi: 10.1017/S0714980816000696. Epub 2017 Jan 10. PMID: 28069090.
39. Fryklund L, Ritzén M, Bertilsson G, Arnlind MH Is the decision on the use of biosimilar growth hormone based on high quality scientific evidence? - a systematic review. Eur J Clin Pharmacol. 2014 May;70(5):509-17. doi: 10.1007/s00228-014-1655-4. Epub 2014 Feb 26.PMID: 24569841 Review.
40. Matar P, Biosimilarity is not a transitive property: implication for interchangeability, naming and pharmacovigilance. Generics and Biosimilars Initiative Journal (GaBI Journal). 2022;11(1):36-40.
DOI: 10.5639/gabij.2022.1101.006
41. Similar medicinal products containing somatropin (Annex to guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues) - Scientific guideline | European Medicines Agency ( June 2018 EMEA/CHMP/BMWP/94528/2005 Rev. 1 Committee for Medicinal Products for Human Use (CHMP) Annex to Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: non-clinical and clinical issues Guideline on similar medicinal products containing somatropin.
42. Graves JA. Utility of the National Cooperative Growth Study database for safety reporting J Pediatr. 1996 May;128(5 Pt 2):S1-3. doi: 10.1016/s0022-3476(96)70001-x.
43. Maghnie M, Ranke MB , Geffner ME , Vlachopapadopoulou E, Ibáñez L , Carlsson M, Cutfield W, Rooman R, Gomez R, Wajnrajch MP, Linglart A , Stawerska R, Clayton PE , Darendeliler F, Hokken-Koelega ACS, Reiko Horikawa R, Tanaka T, Dörr HG , Albertsson-Wikland K , Polak M, Grimberg A. Safety and Efficacy of Pediatric Growth Hormone Therapy: Results From the Full KIGS Cohort. J Clin Endocrinol Metab, 2022 Nov 25;107(12):3287-3301. doi: 10.1210/clinem/dgac517.
44. Blethen SL, MacGillivray MHA risk-benefit assessment of growth hormone use in children Drug Saf. 1997 Nov;17(5):303-16. doi: 10.2165/00002018-199717050-00003.
45. Mergulhão FJ, Taipa MA, Cabral JM, Monteiro GA. Evaluation of bottlenecks in proinsulin secretion by Escherichia coli. J Biotechnol. 2004 Apr 8;109(1-2):31-43. doi: 10.1016/j.jbiotec.2003.10.024. PMID: 15063612.
46. Siebenhofer A, Plank J, Berghold A, Narath M, Gfrerer R, Pieber TR.Short acting insulin analogues versus regular human insulin in patients with diabetes mellitus. Cochrane Database Syst Rev. 2004 Oct 18;(4):CD003287. doi: 10.1002/14651858.CD003287.pub3.
47. Nicolucci A, Ceriello A, Di Bartolo P, Corcos A, Orsini Federici M. Rapid-Acting Insulin Analogues Versus Regular Human Insulin: A Meta-Analysis of Effects on Glycemic Control in Patients with Diabetes. Diabetes Ther. 2020 Mar;11(3):573-584. doi: 10.1007/s13300-019-00732-w. Epub 2019 Dec 23. PMID: 31873857; PMCID: PMC7048883.
48. Fullerton B, Siebenhofer A, Jeitler K, Horvath K, Semlitsch T, Berghold A, Plank J, Thomas R Pieber, Ferdinand M GerlachShort-acting insulin analogues versus regular human insulin for adults with type 1 diabetes mellitus Cochrane Database Syst Rev. 2016 Jun 30;2016(6):CD012161. doi: 10.1002/14651858.CD012161.
49. Monami M , Marchionni N, Mannucci E, Long-acting insulin analogues vs. NPH human insulin in type 1 diabetes. A meta-analysis Diabetes Obes Metab. 2009 Apr;11(4):372-8. doi: 10.1111/j.1463-1326.2008.00976.x.
50. Rossetti P, Porcellati F, Fanelli CG, Perriello G, Torlone E, Bolli GB Superiority of insulin analogues versus human insulin in the treatment of diabetes mellitus Arch Physiol Biochem. 2008 Feb;114(1):3-10. doi: 10.1080/13813450801900777.
51. Tibaldi JM Evolution of insulin: from human to analog Am J Med. 2014 Oct;127(10 Suppl):S25-38. doi: 10.1016/j.amjmed.2014.07.005.
52. Toledano Y, Hadar E , Hod M, Safety of insulin analogues as compared with human insulin in pregnancy Expert Opin Drug Saf. 2016 Jul;15(7):963-73. doi: 10.1080/14740338.2016.1182153. Epub 2016 May 17.
53. Dolinar R, Lavernia F, Edelman S. A Guide To Follow-On Biologics And Biosimilars With A Focus On Insulin. Endocr Pract. 2018 Feb;24(2):195-204. doi: 10.4158/EP161728.RA. PMID: 29466056.
54. Franzè S, Cilurzo F, Minghetti P. Insulin biosimilars: the impact on rapid-acting analogue-based therapy. BioDrugs. 2015 Apr;29(2):113-21. doi: 10.1007/s40259-015-0121-x. PMID: 25773234.
55. Peters AL, Pollom RD, Zielonka JS, Carey MA, Edelman SV. Biosimilars And New Insulin Versions. Endocr Pract. 2015 Dec;21(12):1387-94. doi: 10.4158/EP14595.RA. Epub 2015 Sep 4. PMID: 26340139
56. Rodbard HW, Rodbard D. Biosynthetic Human Insulin and Insulin Analogs Am J Ther. 2020 Jan/Feb;27(1):e42-e51 doi:10.1097/MJT.0000000000001089.
57. Mayer JP, Zhang F, DiMarchi RD Insulin structure and function.Biopolymers. 2007;88(5):687-713. doi: 10.1002/bip.20734.PMID: 17410596 Review.
58. Monnier L, Colette C, Owens D. Basal insulin analogs: from pathophysiology to therapy. What we see, know, and try to comprehend? Diabetes Metab. 2013 Dec;39(6):468-76. doi: 10.1016/j.diabet.2013.09.003. Epub 2013 Oct 17. PMID: 24139826.
59. Yuen KCJ, Miller BS, Boguszewski CL, Hoffman AR. Usefulness and Potential Pitfalls of Long-Acting Growth Hormone Analogs. Front Endocrinol (Lausanne). 2021 Feb 24;12:637209. doi: 10.3389/fendo.2021.637209. Erratum in: Front Endocrinol (Lausanne). 2021 Jun 28;12:705241. PMID: 33716988; PMCID: PMC7943875.
60. Højby Rasmussen M, Janukonyté J , Klose M, Marina D, Tanvig M , Nielsen LF , Höybye C, Andersen M , Feldt-Rasmussen U , Sandahl Christiansen J. Reversible Albumin-Binding GH Possesses a Potential Once-Weekly Treatment Profile in Adult Growth Hormone Deficiency. J Clin Endocrinol Metab. 2016 Mar;101(3):988-98. doi: 10.1210/jc.2015-1991. Epub 2016 Jan 4.
61. Helleberg H, Lindecrona RH, Thygesen P, Bjelke M. Structure identification of circulating metabolites from somapacitan, a long-acting growth hormone derivative, and pharmacokinetics after single and multiple subcutaneous dosing in rats. Eur J Pharm Sci. 2022 Jan 1;168:106032. doi: 10.1016/j.ejps.2021.106032. Epub 2021 Oct 2. PMID: 34610450
62. Thygesen P, Andersen HS, Behrens C, Fels JJ, Nørskov-Lauritsen L, Rischel C, Johansen NL. Nonclinical pharmacokinetic and pharmacodynamic characterisation of somapacitan: A reversible non-covalent albumin-binding growth hormone. Growth Horm IGF Res. 2017 Aug;35:8-16. doi: 10.1016/j.ghir.2017.05.006. Epub 2017 May 24. PMID: 28595133.
63. Miller BS, Blair JC, Rasmussen MH, Maniatis A, Kildemoes RJ, Mori J, Polak M, Bang RB, Böttcher V, Stagi S, Horikawa R. Weekly Somapacitan is Effective and Well Tolerated in Children With GH Deficiency: The Randomized Phase 3 REAL4 Trial. J Clin Endocrinol Metab. 2022 Nov 25;107(12):3378-3388. doi: 10.1210/clinem/dgac513. PMID: 36062966; PMCID: PMC9693810.
64. Kjaer ASL, Jensen RB, Petersen JH, Linneberg A, Kårhus LL, Henriksen LS, Johannsen TH, Main KM, Hoffman AR, Juul A. Tracking and Cumulative Lifetime Exposure to IGF-I in 6459 Healthy Individuals and in SGA Children Treated With GH. J Clin Endocrinol Metab. 2023 Feb 15;108(3):642-652. doi: 10.1210/clinem/dgac605. PMID: 36250350.
65. van der Kaay DCM, Rochtus A, Binder G, Kurth I, Prawitt D, Netchine I, Johannsson G, Hokken-Koelega ACS, Elbracht M, Eggermann T. Comprehensive genetic testing approaches as the basis for personalized management of growth disturbances: current status and perspectives. Endocr Connect. 2022 Oct 10;11(11):e220277. doi: 10.1530/EC-22-0277. PMID: 36064195; PMCID: PMC9578069.
66. de Graaff LC, Clark AJ, Tauber M, Ranke MB, Johnston LB, Caliebe J, Molinas C, Amin N, van Duijn C, Wollmann H, Wallaschofski H, Savage MO, Hokken-Koelega AC. Association analysis of ten candidate genes in a large multinational cohort of small for gestational age children and children with idiopathic short stature (NESTEGG study). Horm Res Paediatr. 2013;80(6):466-76. doi: 10.1159/000355409. Epub 2013 Nov 23. PMID: 24280783.
67. Binder G. Isolated growth hormone deficiency and the GH-1 gene: update 2002. Horm Res. 2002;58 Suppl 3:2-6. doi: 10.1159/000066476. PMID: 12435888
68. Johnston LB, Ester W, Caliebe J, Molinas C, Wollmann H, Fryklund L, Clark AJ, Ranke MB, Tauber M, Hokken Koelega A, Savage MO. Network of European studies of genes in growth (NESTEGG). Horm Res. 2009 Apr;71 Suppl 2:48-54. doi: 10.1159/000192436. Epub 2009 Apr 29. PMID: 19407497.