The development of Interleukin-6 specific peptide antagonists
The development of Interleukin-6 specific peptide antagonists
Eight peptide sequences corresponding to the sites of interaction between IL-6, IL-6 receptor and the signal gp130 were selected by review of the literature. These peptides were synthesized using T-BOC chemistry, purified by HPLC and analyzed for correct molecular weight by Mass Spectroscopy. Each peptide was also used to generate rabbit polyclonal antisera. The effects of each of the eight peptides on the interactions between IL-6, IL-6 receptor and gp130 were studied using three different techniques. These included direct study using biomolecular interaction analysis (BIA) based on plasmon surface resonance and two bioassays based on prostate carcinoma cell line LNCaP proliferation and the aromatase enzyme activity of primary cultures of adipose stromal cells. Binding of the anti-peptide antisera to IL-6 and IL-6 receptor and the effects of the antisera on LNCaP cell proliferation were also studied. In biomolecular interaction studies the IL-6 receptor derived peptide C20E prevented binding of IL-6 to immobilized IL-6R by at least 23% at a 1000-fold excess over IL-6. This peptide also inhibited proliferation of the LNCaP cell line by 24% at 10-5 M and caused decreases in the endogenous aromatase activity of adipose stromal cells of up to 63%. The two peptides G19E and L18S which included overlapping regions of IL-6 involved in IL-6 receptor binding, both bound to immobilized IL-6R in BIA studies. However neither of these two peptides inhibited LNCaP cell proliferation. Peptide G19E caused an increase in adipose stromal cell aromatase activity while the predominant effect of L18S was one of inhibition. Peptide S16M included residues from one site of IL-6 involved in IL-6 receptor binding. This peptide did not bind to IL-6sR in BIA studies, however it did cause significant inhibition of the aromatase activity of adipose stromal cells.
University of Southampton
1999
Bailey, Louise Lyn
(1999)
The development of Interleukin-6 specific peptide antagonists.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Eight peptide sequences corresponding to the sites of interaction between IL-6, IL-6 receptor and the signal gp130 were selected by review of the literature. These peptides were synthesized using T-BOC chemistry, purified by HPLC and analyzed for correct molecular weight by Mass Spectroscopy. Each peptide was also used to generate rabbit polyclonal antisera. The effects of each of the eight peptides on the interactions between IL-6, IL-6 receptor and gp130 were studied using three different techniques. These included direct study using biomolecular interaction analysis (BIA) based on plasmon surface resonance and two bioassays based on prostate carcinoma cell line LNCaP proliferation and the aromatase enzyme activity of primary cultures of adipose stromal cells. Binding of the anti-peptide antisera to IL-6 and IL-6 receptor and the effects of the antisera on LNCaP cell proliferation were also studied. In biomolecular interaction studies the IL-6 receptor derived peptide C20E prevented binding of IL-6 to immobilized IL-6R by at least 23% at a 1000-fold excess over IL-6. This peptide also inhibited proliferation of the LNCaP cell line by 24% at 10-5 M and caused decreases in the endogenous aromatase activity of adipose stromal cells of up to 63%. The two peptides G19E and L18S which included overlapping regions of IL-6 involved in IL-6 receptor binding, both bound to immobilized IL-6R in BIA studies. However neither of these two peptides inhibited LNCaP cell proliferation. Peptide G19E caused an increase in adipose stromal cell aromatase activity while the predominant effect of L18S was one of inhibition. Peptide S16M included residues from one site of IL-6 involved in IL-6 receptor binding. This peptide did not bind to IL-6sR in BIA studies, however it did cause significant inhibition of the aromatase activity of adipose stromal cells.
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Published date: 1999
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Local EPrints ID: 463919
URI: http://eprints.soton.ac.uk/id/eprint/463919
PURE UUID: a6fd8785-127a-43ba-a4d5-2bc3c19850ea
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Date deposited: 04 Jul 2022 20:58
Last modified: 04 Jul 2022 20:58
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Author:
Louise Lyn Bailey
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