【摘要】：目的 3種方法分析10個(gè)血清型肺炎鏈球菌莢膜多糖(pneumococcal capsular polysaccharides,Pn Ps)樣品的分子大小及分子量,并對(duì)這3種方法進(jìn)行比較。方法采用Sepharose CL-4B凝膠排阻層析(SEC)技術(shù)檢測(cè)1型、2型、4型、8型、9N型、12F型、14型、18C型、19A型和23F型Pn Ps各2~3批次樣品在色譜柱中的分配系數(shù)(KD)和樣品組分特征;采用高效凝膠排阻色譜-示差折光檢測(cè)器(high performance size-exclusion chromatography with refractive index,HPSEC-RI)法建立Pullulan標(biāo)準(zhǔn)品分子量標(biāo)準(zhǔn)曲線,計(jì)算各Pn Ps樣品的重均分子量(weight-average molecular weight,Mw);采用高效凝膠排阻色譜-多角度激光光散射儀/示差折光檢測(cè)器聯(lián)用技術(shù)(high performance size-exclu-sion chromatography with multi-angle laser light scattering and refractive index,HPSEC-MALLS/RI)檢測(cè)各Pn Ps樣品的Mw、數(shù)均分子量(number-average molecular weight,Mn)、多分散水平(Mw/Mn)及均方根旋轉(zhuǎn)半徑(root mean square ratio of gyration,Rg);對(duì)HPSEC-RI法和HPSEC-MALLS/RI法檢測(cè)Pullulan分子量標(biāo)準(zhǔn)品的結(jié)果進(jìn)行比較。結(jié)果 SEC技術(shù)不同化學(xué)方法檢測(cè)的同批次Pn Ps樣品的KD值十分接近,且主峰形態(tài)也比較近似,個(gè)別批次Pn Ps不同化學(xué)方法獲得的峰形有差別。經(jīng)HPSEC-RI檢測(cè)Pullulan分子量標(biāo)準(zhǔn)品獲得的回歸方程為y=-0.278 x+10.13,r2=0.998,各Pn Ps樣品的分子量為1.812×105~1.246×106。HPSEC-MALLS/RI聯(lián)用技術(shù)檢測(cè)的各批Pn Ps樣品的Mw分布于9.898×105~1.471×105,Rg為84.9~25.4 nm,同一血清型不同批次樣品Rg間的大小關(guān)系與其Mw間的大小關(guān)系具有較好一致性。結(jié)論結(jié)合多種方法檢測(cè)多糖樣品的分子大小和分子量,可更客觀、全面地了解Pn Ps分子的各種性質(zhì)。
[Abstract]:Objective to analyze the molecular size and molecular weight of 10 serotype Streptococcus pneumoniae capsule polysaccharide (pneumococcal capsular polysaccharides,Pn Ps) samples by three methods and compare them. Methods Sepharose CL-4B gel exclusion chromatography (SEC) technique was used to detect the distribution coefficient (KD) and the sample composition characteristics of 3 batches of type 1, type 2, type 4, type 1, type 9, N, type 1, type 12F, type 14, type 1, type 18C, type 2, Pn Ps, and type 23 F, respectively, in the chromatographic column. The molecular weight standard curve of Pullulan standard product was established by high performance gel exclusion chromatography-differential refraction detector (high performance size-exclusion chromatography with refractive index,HPSEC-RI). The weight average molecular weight (weight-average molecular weight,Mw) of each Pn Ps sample was calculated and the Mw, number of each Pn Ps sample was detected by high performance gel exclusion chromatography-multi-angle laser light scattering spectrometer / differential refractive detector (high performance size-exclu-sion chromatography with multi-angle laser light scattering and refractive index,HPSEC-MALLS/RI). Average molecular weight (number-average molecular weight,Mn), polydispersing level (Mw/Mn) and root-mean-square rotation radius (root mean square ratio of gyration,Rg) were compared between HPSEC-RI and HPSEC-MALLS/RI methods for the detection of Pullulan molecular weight standard. Results the KD values of the same batch Pn Ps samples detected by different chemical methods by SEC technique were very close, and the main peak morphology was similar, and the peak shapes obtained by different chemical methods of individual batches of Pn Ps were different. The regression equation obtained by HPSEC-RI detection of Pullulan molecular weight standard product is yang-0.278x 10.13r2r2O0.998.The molecular weight of each Pn Ps sample is 1.812 脳 105N1.246 脳 106.HPSEC-MALLS/RI. The Mw distribution of each batch of Pn Ps samples is 9.898 脳 105N1.471 脳 105rg = 84.925.4 nm,. The Mw of different batches of samples with the same serotype is larger than that of Rg samples of the same serotype. There is a good consistency between the small relation and the size relation of Mw. Conclusion the molecular size and molecular weight of polysaccharide samples can be detected by various methods, and the properties of Pn Ps molecules can be understood more objectively and comprehensively.
【作者單位】： 蘭州生物制品研究所有限責(zé)任公司第三研究室甘肅省疫苗工程技術(shù)研究中心;
【分類號(hào)】：R378

【參考文獻(xiàn)】

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1 柴雁菁;左智潔;馬波;向左云;施z，

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