Open AccessScreening of lectins from South American plants used as affinity ligands to purify rhEPO
Author(s) -
Gabriel Ignacio Amadeo,
Renato de Azevedo Moreira,
Ricardo José Pontes Lima,
Daniel Câmara Teixeira,
Ricardo Kratje,
Marina Etcheverrigaray
Publication year - 2003
Publication title -
brazilian journal of chemical engineering/brazilian journal of chemical engineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.313
H-Index - 52
eISSN - 1678-4383
pISSN - 0104-6632
DOI - 10.1590/s0104-66322003000100005
Subject(s) - lectin , jacalin , hemagglutination , viscum album , chemistry , biochemistry , artocarpus , ricinus , affinity chromatography , erythrina , ricin , canavalia ensiformis , moraceae , galactose , abrus precatorius , gene isoform , biology , toxin , botany , antigen , enzyme , ecology , gene , genetics
Two groups of isoforms of rhEPO, at a concentration of 300 µg/ml, were tested as putative inhibitors of the lectinic hemagglutination reaction in order to obtain affinity ligand(s) for hormone purification: groups I (pI: 3.80; 3.89; 3.95; 4.07, 4.15 and 4.26) and groups II (pI: 4.15, 4.26; 4.38; 4.51; 4.72 and 4.93) Crude extracts from the vegetable materials Abrus precatorious (Abrin), Artocarpus incisa (Frutalin), Artocarpus integrifolia (Jacalin), Canavalia ensiformes (ConA), Canavalia brasiliensis (Conbr), Cratylia floribunda, Dioclea altissima (DAL), Dioclea grandiflora (DGL), Erythrina vellutina (EVL), Erythrina cristagalli, Lutaelburgia auriculata (lectin not fully characterized yet), Lycopersicum esculentum (LEA), Phaseolus vulgaris (PHA), Ricinus communis (Ricin) and Triticum vulgaris (WGA) were used. Only some of the galactose-specific lectins and the GlcNAc-specific lectins showed rapid full inhibition of the hemagglutination reaction for the less acidic isoforms and the total isoforms of rhEPO, respectively. On this basis, the selected lectins were purified by affinity chromatoghraphy and covalently coupled to cyanogen bromide activated Sepharose® (Amersham-Pharmacia). CHO.K1 cell culture supernatant containing rhEPO was loaded onto the lectin resins and the recoveries were calculated by using specific elutions