GENERATION OF ANTISERUM TO PDGF SYNTHETIC PEPTIDES
GENERATION OF ANTISERUM TO PDGF SYNTHETIC PEPTIDES
批准号:
3425103
负责人:
DANA T GRAVES
金额:
$2.13万
依托单位国家:
美国
项目类别:
财政年份:
1986
资助国家:
美国
项目状态:
已结题
起止时间:
1986-03-01 至 1987-08-31
中文摘要
血小板源性生长因子PDGF是一种多肽激素,
引发涉及伤口愈合、动脉粥样硬化和
恶性肿瘤的发展。 受PDGF影响的细胞内事件包括
胶原和前列腺素合成,癌基因表达,
磷脂酰肌醇周转,酪氨酸磷酸化和细胞内
钙水平。 目前的研究涉及促有丝分裂和趋化
骨肉瘤和血管细胞产生的PDGF样蛋白的作用。
为了进行这些研究,获得一份
识别生物活性的确定区域的抗血清,
未还原的PDGF。 由于纯化PDGF的量不实际,
需要开发抗PDGF血清,提出了一种替代方法
这里. 基于PDGF的氨基酸序列的合成肽将是
合成了 这些肽将被构建,使得第二肽将被构建。
结构通过链内二硫键稳定。 这种方法有
已成功地用于开发一种抗血清,
合成肽和天然B型肝炎抗原。 我们建议
合成四种不同的PDGF肽,将其注射到兔子体内
以产生多克隆抗血清。 然后检测兔抗血清
用于识别合成肽和天然PDGF。
英文摘要
Platelet-derived growth factor, PDGF, is a polypeptide hormone capable of
initiating processes involved in wound healing, atherosclerosis and the
development of malignancy. Intracellular events influenced by PDGF include
collagen and prostaglandin synthesis, oncogene expression,
phosphatidylinositol turnover, tyrosine phosphorylation and intracellular
calcium levels. Studies currently involve the mitogenic and chemotactic
effects of PDGF-like proteins produced by osteosarcoma and vascular cells.
In order to pursue these studies it would be extremely useful to obtain an
antiserum which recognizes a defined region of biologically active,
unreduced PDGF. Since it is not practical to purify PDGF in the quantity
needed to develop an anti-PDGF serum, an alternative approach is presented
here. Synthetic peptides based on the amino acid sequence of PDGF will be
synthesized. These peptides will be constructed so that the secondary
structure is stabilized by intrachain disulfide bridges. This approach has
been successfully used to develop an antiserum which cross reacts with a
synthetic peptide and a native Hepatitis B antigen. We propose to
synthesize four distinct PDGF peptides, which will be injected into rabbits
to generate a polyclonal antiserum. Rabbit antiserum will then be tested
for recognition of the synthetic peptide and native PDGF.
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