Human recombinant antibody fragments specific for a rye-grass pollen allergen: characterization and potential applications

详细信息    查看全文

• 作者：De Lalla ; Claudia ; Tamborini ; Elena ; Longhi ; Renato ; Tresoldi ; Eleonora ; Manoni ; Marco ; Siccardi ; Antonio G. ; et. al.
• 关键词：allergen ; anti-Lol pII ; phage library ; human Fab
• 刊名：Molecular Immunology
• 出版年：1996
• 出版时间：September, 1996
• 年：1996
• 卷：33
• 期：13
• 页码：1049-1058
• 全文大小：1.04 M

文摘

One of the major allergens from the pollen of perennial rye grass (Lolium perenne), Lol pII, was used to isolate specific antibody fragments from a random combinatorial library displaying a large repertoire of human Fab on filamentous phages. After five panning cycles on recombinantLol p II immunotubes, phage binders were isolated and the antibody fragments expressed as soluble Fab molecules in the Escherichia coli periplasm. The DNA sequencing of the clones producing antibodies with the highest binding activity showed three of them to be identical, while one differed by two amino acid substitutions in the heavy chain. The antibody fragments were produced in milligram amounts, affinity-purified and further characterized. They bound the natural allergen as well as the recombinant one, with no cross-reactivity with other allergens contained in the pollen extract of L. perenne. One antibody bound the allergen with K_d