ASSEMBLY OF AN ANTIBODY AND ITS DERIVED ANTIBODY FRAGMENT IN NICOTIANA AND ARABIDOPSIS

被引:117
作者
DENEVE, M
DELOOSE, M
JACOBS, A
VANHOUDT, H
KALUZA, B
WEIDLE, U
VANMONTAGU, M
DEPICKER, A
机构
[1] STATE UNIV GHENT, GENET LAB, KL LEDEGANCKSTR 35, B-9000 GHENT, BELGIUM
[2] BOEHRINGER MANNHEIM GMBH, W-8122 PENZBERG, GERMANY
关键词
CAMV 35S PROMOTER; F(AB) FRAGMENT; INTER-TRANSFORMANT VARIABILITY; MONOCLONAL ANTIBODY; SELF-ASSEMBLY; TRANSGENIC PLANTS; ARABIDOPSIS-THALIANA; NICOTIANA-TABACUM;
D O I
10.1007/BF01977353
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The yield and assembly of an IgG1 antibody and its derived F(ab) fragment were compared in Nicotiana and Arabidopsis. The results obtained showed a lot of interclonal variability. For 45% of the primary transgenic calluses, antigen-binding entities represented less than 0. 1% of the total soluble protein (TSP). Only two of the 103 analysed transformants contained more than 1% of antigen-binding protein, with 1.26% being the highest yield. Analogous amounts of complete antibody and F(ab) accumulated in primary callus tissue. Moreover, yields were in the same range for both species as far as primary callus tissue is concerned. However, the accumulation of the F(ab) fragment in leaf tissue of regenerated plants differed significantly between Nicotiana and Arabidopsis. The F(ab) fragment accumulated to only 0.044% of TSP in Nicotiana leaves but up to 1.3% in Arabidopsis leaves. Furthermore, both species showed differences in the assembly pattern of the complete antibody. Whereas Arabidopsis contained primarily fully assembled antibodies of 150 kDa, Nicotiana showed an abundance of fragments in the 50 kDa range.
引用
收藏
页码:227 / 237
页数:11
相关论文
共 31 条
  • [1] 2-DIMENSIONAL GEL-ELECTROPHORESIS, PROTEIN ELECTROBLOTTING AND MICROSEQUENCING - A DIRECT LINK BETWEEN PROTEINS AND GENES
    BAUW, G
    RASMUSSEN, HH
    VANDENBULCKE, M
    VANDAMME, J
    PUYPE, M
    GESSER, B
    CELIS, JE
    VANDERKERCKHOVE, J
    [J]. ELECTROPHORESIS, 1990, 11 (07) : 528 - 536
  • [2] PHYTOANTIBODIES - A GENERAL VECTOR FOR THE EXPRESSION OF IMMUNOGLOBULIN DOMAINS IN TRANSGENIC PLANTS
    BENVENUTO, E
    ORDAS, RJ
    TAVAZZA, R
    ANCORA, G
    BIOCCA, S
    CATTANEO, A
    GALEFFI, P
    [J]. PLANT MOLECULAR BIOLOGY, 1991, 17 (04) : 865 - 874
  • [3] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
  • [4] EFFECT OF T-DNA CONFIGURATION ON TRANSGENE EXPRESSION
    BREYNE, P
    GHEYSEN, G
    JACOBS, A
    VANMONTAGU, M
    DEPICKER, A
    [J]. MOLECULAR & GENERAL GENETICS, 1992, 235 (2-3): : 389 - 396
  • [5] RENATURATION, PURIFICATION AND CHARACTERIZATION OF RECOMBINANT FAB-FRAGMENTS PRODUCED IN ESCHERICHIA-COLI
    BUCHNER, J
    RUDOLPH, R
    [J]. BIO-TECHNOLOGY, 1991, 9 (02): : 157 - 162
  • [6] CLONING AND NUCLEOTIDE-SEQUENCE OF HEAVY-CHAIN AND LIGHT-CHAIN CDNAS FROM A CREATINE-KINASE-SPECIFIC MONOCLONAL-ANTIBODY
    BUCKEL, P
    HUBNERPARAJSZ, C
    MATTES, R
    LENZ, H
    HAUG, H
    BEAUCAMP, K
    [J]. GENE, 1987, 51 (01) : 13 - 19
  • [7] CHRISPEELS MJ, 1991, ANNU REV PLANT PHYS, V42, P21, DOI 10.1146/annurev.pp.42.060191.000321
  • [8] De Greve H, 1982, J Mol Appl Genet, V1, P499
  • [9] DEBLAERE R, 1985, NUCLEIC ACIDS RES, V13, P4777, DOI 10.1093/nar/13.13.4777
  • [10] PLANT AND MAMMALIAN SORTING SIGNALS FOR PROTEIN RETENTION IN THE ENDOPLASMIC-RETICULUM CONTAIN A CONSERVED EPITOPE
    DENECKE, J
    DERYCKE, R
    BOTTERMAN, J
    [J]. EMBO JOURNAL, 1992, 11 (06) : 2345 - 2355