* Senior Investigator: Joseph Shiloach

* Affiliation: Biotechnology Unit, LCDB/NIDDK.

* Research: Optimizing growth, production, and product recovery processes involving prokaryotes and eukaryotes, both native and recombinant. Concentrating on protein production and purification, with an emphasis on scale-up.

* Personnel/Resources: B. Kaufman (microbiologist), A. Tsai (protein purification).

The unit has a modern state-of-the-art pilot production plant with a capability of processing bacterial culture up to 300 liters and mammalian cell culture up to 40 liters. The facility has the ability to perform large scale production and purification of various biological products for various purposes such as clinical experiments and structural studies.

* Publications:

S. Reuveny, Y.J. Kim, C.W. Kemp, & J. Shiloach (1993). Biotechnology and Bioengineering, 42: 235-239. Production of recombinant proteins in high-density insect cell cultures.

M. DiBrino, K.C. Parker, J. Shiloach, M. Knierman, J. Lukszo, R.V. Turner, W.E. Biddison, & J. E. Coligan (1993). Proc Natl Acad Sci. USA 4:1508-12. Endogenous peptides bound to HLA-A3 possess a specific combination of anchor residues that permit identification of potential antigenic peptides.

A. Fattom, R. Schneerson, D.C. Watson, W.W. Karakawa, D. Fitzgerald, I. Pastan, X. Li, J. Shiloach, D.A. Bryla, & J. B. Robbins (1993). Infect Immun 3:1023-32. Laboratory and clinical evaluation of conjugate vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides bound to Pseudomonas aeruginosa recombinant exoprotein A.

J. Hsiao, M. Ahluwalia, J.B. Kaufman, T.R. Clem, & J. Shiloach. (1992). Ann NY Acad Sci. 665:320-33. Adaptive control strategy for maintaining dissolved oxygen concentration in high density growth of recombinant E. coli.

S. Reuveny, C.W. Kemp, L. Eppstein, & J. Shiloach (1992). Ann NY Acad Sci. 665:230-7. Carbohydrate metabolism in insect cell cultures during cell growth and recombinant protein production.

S.P. Becerra, A. Kumar, M.S. Lewis, S.G. Widen, J. Abbotts, E. M. Karawya, S.H. Hughes, J. Shiloach, S.H. Wilson, & M. S. Lewis (1991). Biochemistry 50:11707-19. Protein-protein interactions of HIV-1 reverse transcriptase: implication of central and C-terminal regions in subunit binding.

* Contact: Dr. J. Shiloach, Bldg. 6, Room B1-33. NIDDK, NIH, Bethesda, MD 20892.

Tel: (301) 496-9719 Fax: (301) 496-5239

Figure: Recombinant exotoxin A purification.