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背景知识


凝集素(Lectins)是动物细胞和植物细胞都能够合成和分泌的、能与糖结合的蛋白质。因其能凝集红血球(含血型物质),故名凝集素。如今,凝集素指所有糖结合蛋白,不管其是否能凝集红细胞。凝集素能识别糖蛋白和糖肽中,特别是细胞膜中复杂的碳水化合物结构,即细胞膜表面的碳脂化合物决定簇。一种凝集素具有对某一种特异性糖基专一性结合的能力。凝集素在生物识别现象中发挥作用,涉及细胞和蛋白的识别,比如:一些病毒利用凝集素将其附着在宿主细胞上。目前,已经在植物、病毒、微生物和动物中发现了凝集素,但许多生物学功能仍待探究。


大多数凝集素为多聚体,由非共价连接的亚基组成。这种多聚化赋予了凝集素凝集细胞和与糖结合物形成沉淀的能力(类似于抗原-抗体反应)。凝集素与糖结构的结合具有高度的特异性。凝集素与其受体的结合亲和力因其受体糖结构的细微变化而有较大差异。

 

〈〈 凝集素在动物中的作用

凝集素在动物中发挥不同的生物学功能,从细胞黏附到糖蛋白合成的调控,以及血液中蛋白水平的调控。

也可与胞外及胞内的可溶性糖蛋白结合。有些凝集素存在于哺乳动物肝细胞表面,可特异性识别半乳糖(galactose)残基。这些表面受体的作用在于从循环系统中清除某些糖蛋白。另外一凝集素也是一个受体,可识别含甘露糖-6-磷酸的水解酶(mannose-6-phosphate), 而将靶蛋白转入溶酶体。

已知,凝集素在免疫系统中发挥重要重用,可识别广泛存在于病原体上的糖。例如凝集素补体活化途径(complement activation pathway )和甘露糖结合凝集素(mannose-binding lectin)。

 

〈〈 凝集素在植物中的作用

凝集素在植物 (豆科凝集素,Legume lectins) 中的功能仍不确切。植物种子中的凝集素随植物生长,其浓度大量降低,表明凝集素在植物种子萌发及种子存活方面发挥重要作用。

研究发现,许多植物凝集素识别的并非糖配体,这些配体主要以疏水性配体为主包括腺嘌呤、植物激素、细胞分裂素、吲哚乙酸,也包括水溶性的植物激素。

凝集素在凝集红细胞方面与抗体有类似的能力,但凝集素并非免疫细胞的组分。

 

〈〈 凝集素纯化方法

 传统方法

盐诱导结晶(salt-induced crystallization)、乙醇沉淀(ethanol  precipitation)、离子交换层析(Ion exchange chromatography)、凝胶过滤

 亲和层析

(affinity chromatography)

 

利用凝集素和糖结构的特异性结合特性


Vectorlabs公司既采用传统方法也采用亲和层析方法。产品经多种方法验证,如抗血清免疫沉淀、凝集滴度、多聚酰胺凝胶电泳、特异性亲和柱 结合活性分析等。

 

凝集素结合特异性指南

 糖

 凝集素

 Fucose(Fuc)

 AAL,LTL, UEA I

 Galactose(GalNAc) 

 ACL, ECL, EEL, GSL, GSL I-B4, Jacalin, MAL I, PNA, RCA I,  RCA II, SBA

 Glucose(Glc)

 Con A, LCA, PSA

 Mannose(Man) 

 Con A, GNL, HHL, LCA, NPL, PSA

 N-Acetylgalactosamine

 (GalNAc)

 BPL, DBA, GSL I, MPL, PTL, RCA I, RCA II, SJA, SBA, VVA, WFA

 N-Acetylglucosamine(GlcNAc)

 DSL, GSL II,LEL, STL, WGA

 Sialic Acid(SA)

 MAL II, SNA

 Complex Structures

 PHA-E, PHA-L


Vectorlabs供应凝集素类型 

 

〈〈 琼脂糖结合凝集素

凝集素亲和层析是一种简广泛应用的用于分离糖复合物的技术。糖复合物与固定的凝集素结合,未结合的物质被洗掉,再经过糖蛋白洗脱液(抑制凝集素的结合)洗脱即可。

 

通过此方法,可分离纯化可溶性蛋白、激素、抗原、多糖、洗涤剂可溶性膜结合糖复合物,细胞表面受体、血型物质、病毒糖蛋白、组织相容性抗原,淋巴因子、酶、淋巴细胞标记、血清蛋白、胚胎抗原等。

 

Vectorlabs选择亲和纯化的凝集素,采用耐热的、交联度4%的琼脂糖珠作为固相基质,通过共价结合的方式,在凝集素和琼脂糖珠间插入一个亲水间隔臂。这种结合方式可维持凝集素活性,最小化结合凝集素发生的构象变化,避免可能造成的非特异性离子或疏水反应的发生。

 

产品特点

● 最大化保留结合凝集素的碳水化合物结合活性

● 在较广的pH范围内线性稳定

● 与琼脂糖珠结合的蛋白复合物不易被Tris或常规的Buffer洗脱

● 非特异性结合少

 

〈〈 带标记凝集素

某些凝集素经高度复合后仍具有较高的活性,可溶,非特异性结合低,而有些则不能。

Vectorlabs提供的每个带标记的凝集素具有适当数量的荧光基团与或生物素,从而提供了最佳的凝集素染色特性。

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