{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/94481"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/94481","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Isolation of a human milk sialyloligosaccharide by affinity chromatography with wheat germ agglutinin (WGA)","abstract":"Lectin affinity chromatography has been applied to the separation of the sialyloligosaccharides of human milk. A human milk sialyloligosaccharide fraction was tritium labeled and applied to a highly substituted WGA-agarose column (20 mg/ml). Only a single component from the complete sialyloligosaccharide fraction was retarded in the WGA-agarose column. The WGA-bound fraction when applied to paper chromatography migrated with identical mobility as the sialylhexasaccharide fraction (S-5) of human milk, previously isolated and described by Kobata and Ginsburg in 1972 [Arch. Biochem. Biophys., 150:273-281]. A purified sialylhexasaccharide fraction (S-5), isolated according to the method of Kobata and Ginsburg, was radiolabeled and applied to the WGA-agarose column. The WGA-bound (60%) and WGA-unbound (40%) sialylhexasaccharide fractions were isolated. The WGA-bound sialylhexasaccharide fraction was subjected to neuraminidase digestion to remove sialic acid, and the resulting neutral oligosaccharide had more affinity for the WGA-agarose column. Sequential exoglycosidase digestion of the asialo derivative of the WGA-bound fraction with jack bean β -galactosidase and β -hexosaminidase demonstrated the presence of a lacto-N-neohexaose core. The position of sialic acid in the sialyllacto-N-neohexaose was determined by simultaneous digestion of the sialylhexaose with jack bean β -galactosidase and β -hexosaminidase, which removed the non-sialylated branch from the sialylhexaose and produced a sialyltetraose. The sialyltetraose was found to be sialyltetrasaccharide c as demonstrated by its elution time on HPLC and direct binding to monospecific anti-sialyltetrasccharide c serum. The structural data indicated that the WGA-bound sialylhexaose is a sialyl derivative of lacto-N-neohexaose with sialic acid linked to the 3 branch of this core structure which represents a previously undescribed sialyloligosaccharide in human milk. The structure of the WGA-bound sialylhexaose is,","abstract_html":"Lectin affinity chromatography has been applied to the separation of the sialyloligosaccharides of human milk. A human milk sialyloligosaccharide fraction was tritium labeled and applied to a highly substituted WGA-agarose column (20 mg/ml). Only a single component from the complete sialyloligosaccharide fraction was retarded in the WGA-agarose column. The WGA-bound fraction when applied to paper chromatography migrated with identical mobility as the sialylhexasaccharide fraction (S-5) of human milk, previously isolated and described by Kobata and Ginsburg in 1972 [Arch. Biochem. Biophys., 150:273-281]. A purified sialylhexasaccharide fraction (S-5), isolated according to the method of Kobata and Ginsburg, was radiolabeled and applied to the WGA-agarose column. The WGA-bound (60%) and WGA-unbound (40%) sialylhexasaccharide fractions were isolated. The WGA-bound sialylhexasaccharide fraction was subjected to neuraminidase digestion to remove sialic acid, and the resulting neutral oligosaccharide had more affinity for the WGA-agarose column. Sequential exoglycosidase digestion of the asialo derivative of the WGA-bound fraction with jack bean β -galactosidase and β -hexosaminidase demonstrated the presence of a lacto-N-neohexaose core. The position of sialic acid in the sialyllacto-N-neohexaose was determined by simultaneous digestion of the sialylhexaose with jack bean β -galactosidase and β -hexosaminidase, which removed the non-sialylated branch from the sialylhexaose and produced a sialyltetraose. The sialyltetraose was found to be sialyltetrasaccharide c as demonstrated by its elution time on HPLC and direct binding to monospecific anti-sialyltetrasccharide c serum. The structural data indicated that the WGA-bound sialylhexaose is a sialyl derivative of lacto-N-neohexaose with sialic acid linked to the 3 branch of this core structure which represents a previously undescribed sialyloligosaccharide in human milk. The structure of the WGA-bound sialylhexaose is,","abstract_has_math":false,"creators":["Tarrago-Trani, Maria T."],"institution":"Virginia Polytechnic Institute and State University","degree_name":"M.S.","degree_level":"masters","degree_discipline":"Biochemistry","degree_department":"Biochemistry","school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":1986,"date_issued":"1986","date_published":"1986","updated_at":"2026-07-22T22:20:37Z","subjects":[],"languages":["en_US"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10919/94481","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.department","label":"Department","values":["Biochemistry"]},{"key":"dc:creator","label":"Author","values":["Tarrago-Trani, Maria T."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2019-10-10T19:11:57Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2019-10-10T19:11:57Z"]},{"key":"dc:date.issued","label":"Date","values":["1986"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Polytechnic Institute and State University"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.dcmitype","label":"Dc Type Dcmitype","values":["Text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Virginia Polytechnic Institute and State University"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]},{"key":"dc:rights","label":"Dc Rights","values":["In Copyright"]},{"key":"dc:rights.uri","label":"Rights URI","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10919/94481"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Lectin affinity chromatography has been applied to the separation of the sialyloligosaccharides of human milk. A human milk sialyloligosaccharide fraction was tritium labeled and applied to a highly substituted WGA-agarose column (20 mg/ml). Only a single component from the complete sialyloligosaccharide fraction was retarded in the WGA-agarose column. The WGA-bound fraction when applied to paper chromatography migrated with identical mobility as the sialylhexasaccharide fraction (S-5) of human milk, previously isolated and described by Kobata and Ginsburg in 1972 [Arch. Biochem. Biophys., 150:273-281]. A purified sialylhexasaccharide fraction (S-5), isolated according to the method of Kobata and Ginsburg, was radiolabeled and applied to the WGA-agarose column. The WGA-bound (60%) and WGA-unbound (40%) sialylhexasaccharide fractions were isolated. The WGA-bound sialylhexasaccharide fraction was subjected to neuraminidase digestion to remove sialic acid, and the resulting neutral oligosaccharide had more affinity for the WGA-agarose column. Sequential exoglycosidase digestion of the asialo derivative of the WGA-bound fraction with jack bean β -galactosidase and β -hexosaminidase demonstrated the presence of a lacto-N-neohexaose core. The position of sialic acid in the sialyllacto-N-neohexaose was determined by simultaneous digestion of the sialylhexaose with jack bean β -galactosidase and β -hexosaminidase, which removed the non-sialylated branch from the sialylhexaose and produced a sialyltetraose. The sialyltetraose was found to be sialyltetrasaccharide c as demonstrated by its elution time on HPLC and direct binding to monospecific anti-sialyltetrasccharide c serum. The structural data indicated that the WGA-bound sialylhexaose is a sialyl derivative of lacto-N-neohexaose with sialic acid linked to the 3 branch of this core structure which represents a previously undescribed sialyloligosaccharide in human milk. The structure of the WGA-bound sialylhexaose is,"]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["M.S."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Isolation of a human milk sialyloligosaccharide by affinity chromatography with wheat germ agglutinin (WGA)"]}]}],"canonical_facts":{"dc:contributor.department":["Biochemistry"],"dc:creator":["Tarrago-Trani, Maria T."],"dc:date.accessioned":["2019-10-10T19:11:57Z"],"dc:date.available":["2019-10-10T19:11:57Z"],"dc:date.issued":["1986"],"dc:description.abstract":["Lectin affinity chromatography has been applied to the separation of the sialyloligosaccharides of human milk. A human milk sialyloligosaccharide fraction was tritium labeled and applied to a highly substituted WGA-agarose column (20 mg/ml). Only a single component from the complete sialyloligosaccharide fraction was retarded in the WGA-agarose column. The WGA-bound fraction when applied to paper chromatography migrated with identical mobility as the sialylhexasaccharide fraction (S-5) of human milk, previously isolated and described by Kobata and Ginsburg in 1972 [Arch. Biochem. Biophys., 150:273-281]. A purified sialylhexasaccharide fraction (S-5), isolated according to the method of Kobata and Ginsburg, was radiolabeled and applied to the WGA-agarose column. The WGA-bound (60%) and WGA-unbound (40%) sialylhexasaccharide fractions were isolated. The WGA-bound sialylhexasaccharide fraction was subjected to neuraminidase digestion to remove sialic acid, and the resulting neutral oligosaccharide had more affinity for the WGA-agarose column. Sequential exoglycosidase digestion of the asialo derivative of the WGA-bound fraction with jack bean β -galactosidase and β -hexosaminidase demonstrated the presence of a lacto-N-neohexaose core. The position of sialic acid in the sialyllacto-N-neohexaose was determined by simultaneous digestion of the sialylhexaose with jack bean β -galactosidase and β -hexosaminidase, which removed the non-sialylated branch from the sialylhexaose and produced a sialyltetraose. The sialyltetraose was found to be sialyltetrasaccharide c as demonstrated by its elution time on HPLC and direct binding to monospecific anti-sialyltetrasccharide c serum. The structural data indicated that the WGA-bound sialylhexaose is a sialyl derivative of lacto-N-neohexaose with sialic acid linked to the 3 branch of this core structure which represents a previously undescribed sialyloligosaccharide in human milk. The structure of the WGA-bound sialylhexaose is,"],"dc:description.degree":["M.S."],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["http://hdl.handle.net/10919/94481"],"dc:language.iso":["en_US"],"dc:publisher":["Virginia Polytechnic Institute and State University"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:title":["Isolation of a human milk sialyloligosaccharide by affinity chromatography with wheat germ agglutinin (WGA)"],"dc:type":["Thesis"],"dc:type.dcmitype":["Text"],"thesis:degree_discipline":["Biochemistry"],"thesis:degree_level":["masters"],"thesis:degree_name":["M.S."],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:20:37Z"}