Monoclonal antibodies selective for low molecular weight neurofilaments
Monoclonal antibodies selective for low molecular weight neurofilaments
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DOI：
10.1089/15368590252917647
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发表时间：
2002-02-01
期刊：
影响因子：
--
通讯作者：
Stigbrand, T
中科院分区：
文献类型：
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作者：
Norgren, N;Karlsson, JE;Stigbrand, T
神经丝是维持神经细胞轴突直径和结构组织所必需的。神经丝的低分子量形式，即神经丝的核心神经丝轻蛋白作为免疫原，与这种形式的神经丝产生选择性反应性的杂交瘤。在大约100个测试的克隆中，有6个杂交瘤具有高度的歧视性。所有涉及的表位都定位于抗原的杆状区域，这是通过α -糜蛋白酶消化纯化的纤维和酶促肽定位确定的。覆盖整个棒状区域的合成肽（20米）与抗体不发生反应。利用噬菌体展示肽文库鉴定抗体的4个一致序列。结果表明，所有表位均为构象型。BIAcore数据进一步表明，这两个表位是独立的。获得这种特异性试剂是进一步阐明神经丝蛋白低分子量形式生物学的先决条件。
Neurofilaments are necessary for the maintenance of axonal caliber and structural organization of nerve cells. The low molecular weight form of neurofilament, the neurofilament light protein, which serves as the core of the filament, was used as immunogen for generation of hybridomas with selective reactivity with this form of the filament. Six hybridomas, out of approximately 100 tested clones, were highly discriminatory. All involved epitopes were localized to the rod region of the antigen, as determined by alpha-chymotrypsin digestion of the purified filament and enzymatic peptide mapping. Synthetic peptides (20 mers) covering the entire rod region did not react with the antibodies. A phage display peptide library was used to identify four consensus sequences for the antibodies. The results indicate that all epitopes were of conformational type. Pair-wise BIAcore data furthermore indicated that the epitopes were independent. The access to such specific reagents is a prerequisite for further elucidation of the biology of the low molecular weight form of neurofilaments proteins.