Site-Specific Conjugation of Bifunctional Chelator BAT to Mouse IgG1 Fab’ Fragment_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LI Jun , WANG Xuehao , CHENG Feng , WANG Xiaoming , CHEN Zhaolei , SHEN Xiping.

Corresponding author：

Keywords：

Antibody Monoclonal Chelator Conjugation

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

【Abstract】ObjectiveTo introduce a new approach for site-specific conjugation of Fab’ fragments of mouse monoclonal antibody (MoAb) to a bifunctional chelator 6-〔p-(bromoacetamido)benzyl〕-1,4,8,11 -tetraazacyclo-tetradecane-N,N′,N″,N-tetraacetic acid (BAT).
MethodsFab’ fragments of B43 MoAb were obtained by a simple twostep method： 1, stable F(ab’)2 was produced by pepsin treatment of B43 MoAb. 2, Fab’ was obtained by reduction of the F(ab’)2 with cysteine. Fab’ fragments were directly conjugated with BAT.
ResultsFab’ contained about 1.8 free thiol per molecule. The conjugation rate was 74% and each Fab’ fragment carried 1.28 BAT. The F(ab’)2, Fab’ and Fab’-BAT all maintained good biologic activity.
ConclusionThis study may provide a simple and efficient method conjugating of MoAb Fab’ fragments to BAT.

Citation： LI Jun,WANG Xuehao,CHENG Feng,WANG Xiaoming,CHEN Zhaolei,SHEN Xiping.. Site-Specific Conjugation of Bifunctional Chelator BAT to Mouse IgG1 Fab’ Fragment. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2005, 12(3): 268-270. doi: Copy

1.	Kaklamani V, O’Regan RM. New targeted therapies in breast cancer [Ｊ]. Semin Oncol， 2004; 31(2 Suppl 4)∶20.
2.	Sanz L, Blanco B, AlvarezVallina L. Antibodies and gene therapy: teaching old ‘magic bullets’ new tricks [Ｊ]. Trends Immunol, 2004; 25(2)∶85.
3.	Merdan T, Callahan J, Petersen H, et al. Pegylated polyethylenimineFab’ antibody fragment conjugates for targeted gene delivery to human ovarian carcinoma cells [Ｊ]. Bioconjug Chem, 2003; 14(5)∶989.
4.	Von Rothenburg T, Schaffstein J, Ludwig J, et al. Imaging osteomyelitis with Tc99mlabeled antigranulocyte antibody Fab’ fragments [Ｊ]. Clin Nucl Med, 2003; 28(8)∶643.
5.	Liu X, Zhen Y. Antitumor effects of monoclonal antibody Fab’ fragmentcontaining immunoconjugates [Ｊ]. Chin Med Sci J, 2002; 17(1)∶1.
6.	Hashida S, Imagawa M, Inoue S, et al. More useful maleimide compounds for the conjugation of Fab’ to horseradish peroxidase through thiol groups in the hinge [Ｊ]. J Appl Biochem, 1984; 6(1-2)∶56.
7.	Smith A, Alberto R, Blaeuenstein P, et al. Preclinical evaluation of 67Culabeled intact and fragmented anticolon carcinoma monoclonal antibody MAb35 [Ｊ]. Cancer Res, 1993; 53(23)∶5727.
8.	McCall MJ, Diril H, Meares CF. Simplified method for conjugating macrocyclic bifunctional chelating agents to antibodies via 2iminothiolane [Ｊ]. Bioconjug Chem, 1990; 1(3)∶222.
9.	Anderson CJ, Schwarz SW, Connett JM, et al. Preparation, biodistribution and dosimetry of copper64labeled anticolorectal carcinoma monoclonal antibody fragments 1A3F(ab’)2 [Ｊ]. J Nucl Med, 1995; 36(5)∶850.
10.	Riddles PW, Blakeley RL, Zerner B. Reassessment of Ellman’s reagent [Ｊ]. Methods Enzymol, 1983; 91（1）∶49.
11.	Meares CF, McCall MJ, Reardan DT, et al. Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions [Ｊ]. Anal Biochem, 1984; 142(1)∶68.
12.	SmithJones PM, Fridrich R, Kaden TA, et al. Antibody labeling with copper67 using the bifunctional macrocycle 4[(1,4,8,11tetraazacyclotetradec1yl)methyl]benzoic acid [Ｊ]. Bioconjug Chem, 1991; 2(6)∶415.
13.	Parham P, Androlewicz MJ, Brodsky FM, et al. Monoclonal antibodies: purification, fragmentation and application to structural and functional studies of class I MHC antigens [Ｊ]. J Immunol Methods, 1982; 53(2)∶133.

1. Kaklamani V, O’Regan RM. New targeted therapies in breast cancer [Ｊ]. Semin Oncol， 2004; 31(2 Suppl 4)∶20.
2. Sanz L, Blanco B, AlvarezVallina L. Antibodies and gene therapy: teaching old ‘magic bullets’ new tricks [Ｊ]. Trends Immunol, 2004; 25(2)∶85.
3. Merdan T, Callahan J, Petersen H, et al. Pegylated polyethylenimineFab’ antibody fragment conjugates for targeted gene delivery to human ovarian carcinoma cells [Ｊ]. Bioconjug Chem, 2003; 14(5)∶989.
4. Von Rothenburg T, Schaffstein J, Ludwig J, et al. Imaging osteomyelitis with Tc99mlabeled antigranulocyte antibody Fab’ fragments [Ｊ]. Clin Nucl Med, 2003; 28(8)∶643.
5. Liu X, Zhen Y. Antitumor effects of monoclonal antibody Fab’ fragmentcontaining immunoconjugates [Ｊ]. Chin Med Sci J, 2002; 17(1)∶1.
6. Hashida S, Imagawa M, Inoue S, et al. More useful maleimide compounds for the conjugation of Fab’ to horseradish peroxidase through thiol groups in the hinge [Ｊ]. J Appl Biochem, 1984; 6(1-2)∶56.
7. Smith A, Alberto R, Blaeuenstein P, et al. Preclinical evaluation of 67Culabeled intact and fragmented anticolon carcinoma monoclonal antibody MAb35 [Ｊ]. Cancer Res, 1993; 53(23)∶5727.
8. McCall MJ, Diril H, Meares CF. Simplified method for conjugating macrocyclic bifunctional chelating agents to antibodies via 2iminothiolane [Ｊ]. Bioconjug Chem, 1990; 1(3)∶222.
9. Anderson CJ, Schwarz SW, Connett JM, et al. Preparation, biodistribution and dosimetry of copper64labeled anticolorectal carcinoma monoclonal antibody fragments 1A3F(ab’)2 [Ｊ]. J Nucl Med, 1995; 36(5)∶850.
10. Riddles PW, Blakeley RL, Zerner B. Reassessment of Ellman’s reagent [Ｊ]. Methods Enzymol, 1983; 91（1）∶49.
11. Meares CF, McCall MJ, Reardan DT, et al. Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions [Ｊ]. Anal Biochem, 1984; 142(1)∶68.
12. SmithJones PM, Fridrich R, Kaden TA, et al. Antibody labeling with copper67 using the bifunctional macrocycle 4[(1,4,8,11tetraazacyclotetradec1yl)methyl]benzoic acid [Ｊ]. Bioconjug Chem, 1991; 2(6)∶415.
13. Parham P, Androlewicz MJ, Brodsky FM, et al. Monoclonal antibodies: purification, fragmentation and application to structural and functional studies of class I MHC antigens [Ｊ]. J Immunol Methods, 1982; 53(2)∶133.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Site-Specific Conjugation of Bifunctional Chelator BAT to Mouse IgG1 Fab’ Fragment

Abstract Full text Figures/Tables Video References Cited by

Format

Content