Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide.

L Brade; P Kosma; B J Appelmelk; H Paulsen; H Brade

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Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide.

机译：使用合成抗原来确定针对细菌脂多糖的3-deoxy-D-manno-octulosonate区域的单克隆抗体的表位特异性。

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Mouse monoclonal antibodies were raised against heat-killed bacteria of the Re mutant R595 of Salmonella minnesota and characterized by the passive hemolysis and passive hemolysis inhibition tests and by double immunodiffusion experiments using lipopolysaccharide (LPS) from different rough mutants of S. minnesota and synthetic antigens. The latter were copolymerization products of acrylamide with the alpha- and beta-allylglycosides of 3-deoxy-D-manno-octulosonic acid (KDO) and the alpha-2,4-linked KDO disaccharide [poly-alpha-KDO, poly-beta-KDO, and poly-(alpha-KDO)2, respectively], and sodium (3-deoxy-D-manno-octulopyranosyl)onate-(2----6)-(2-deoxy-2-[ (R)-3- hydroxytetradecanoylamino]- beta-D-glucopyranosyl)-(1----6)-(2-deoxy-2-[(R)-3-hydroxytetradecanoy lam ino]-D-glucose) [alpha-KDO-(GlcNhm)2], representing a part structure of Re LPS. One antibody (clone 20, immunoglobulin M) was found to recognize a terminal alpha-linked KDO residue, since it reacted in the passive hemolysis assay with alpha-KDO-(GlcNhm)2 and all LPS tested, it was inhibited by all synthetic antigens containing alpha-linked KDO residues, and it gave a reaction of identity with poly-alpha-KDO and poly-(alpha-KDO)2 in double immunodiffusion experiments. A second antibody (clone 25, immunoglobulin G3) was identified as specific for an alpha-2,4-linked KDO disaccharide, since it reacted in immunodiffusion exclusively with synthetic poly-(alpha-KDO)2 and not with the monosaccharide derivatives in either anomeric configuration, and it was inhibited only with poly-(alpha-KDO)2 and with LPS from S. minnesota R595 (Re) and R345 (Rb2). The reaction of this antibody with R345 LPS is attributed to the quantitative substitution with KDO disaccharide present as a side chain, which is not present in stoichiometric amounts in the other LPS.

机译：产生了针对明尼苏达沙门氏菌Re突变体R595的热灭活细菌的小鼠单克隆抗体，其特征在于被动溶血和被动溶血抑制试验，以及使用来自明尼苏达州不同粗突变体和合成抗原的脂多糖（LPS）进行的双重免疫扩散实验。。后者是丙烯酰胺与3-脱氧-D-甘露糖辛酸（KDO）和α-2,4-连接的KDO二糖[聚-α-KDO，聚-β]的α-和β-烯丙基糖苷的共聚产物。 -KDO和聚（α-KDO）2]和（3-脱氧-D-甘露聚糖-八吡喃糖基）磺酸钠-（2 ---- 6）-（2-脱氧-2- [ ）-3-羟基十四烷酰氨基]-β-D-吡喃葡萄糖基）-（1 ---- 6）-（2-脱氧-2-[（R）-3-羟基十四烷酰氨基] -D-葡萄糖）[alpha-KDO -（GlcNhm）2]，表示Re LPS的零件结构。发现一种抗体（克隆20，免疫球蛋白M）可识别末端的α-连接的KDO残基，因为它在被动溶血试验中与α-KDO-（GlcNhm）2反应，并且测试的所有LPS均被所有合成抗原抑制含有α-连接的KDO残基，并且在双重免疫扩散实验中，它与poly-α-KDO和poly-（α-KDO）2产生了同一性反应。第二种抗体（克隆25，免疫球蛋白G3）被鉴定为对α-2,4-连接的KDO二糖具有特异性，因为它在免疫扩散中仅与合成的聚-（α-KDO）2发生反应，而与任一中的单糖衍生物均不发生反应异构构型，并且仅被聚-α-KDO2和来自明尼苏达州立大学R595（Re）和R345（Rb2）的LPS抑制。该抗体与R345 LPS的反应归因于以侧链存在的KDO二糖的定量取代，而KDO二糖在其他LPS中以化学计量的形式不存在。

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《Infection and immunity》 |1987年第2期|共5页
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L Brade; P Kosma; B J Appelmelk; H Paulsen; H Brade;
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6. Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide. [O] . L Brade, P Kosma, B J Appelmelk, 1987

机译：使用合成抗原来确定针对细菌脂多糖的3-deoxy-D-manno-octulosonate区域的单克隆抗体的表位特异性。
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Use of synthetic antigens to determine the epitope specificities of monoclonal antibodies against the 3-deoxy-D-manno-octulosonate region of bacterial lipopolysaccharide.

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