Formation of multimeric antibodies for self-delivery of active monomers.
Proteins and peptides have been used as drugs for almost a century. Technological advances in the past 30 years have enabled the production of pure, stable proteins in vast amounts. In contrast, administration of proteins based on their native active conformation (and thus necessitating the use of subcutaneous injections) has remained solely unchanged. The therapeutic anti-HER2 humanized monoclonal immunoglobulin (IgG) Trastuzumab (Herceptin) is a first line of the treatment for breast cancer. Chicken IgY is a commercially important polyclonal antibody (Ab). These Abs were examined for their ability to self-assemble and form ordered aggregates, by several biophysical methods. Atomic force microscopy analyses revealed the formation of multimeric nanostructures. The biological activity of multimeric IgG or IgY particles was retained and restored, in a dilution/time-dependent manner. IgG activity was confirmed by a binding assay using HER2 + human breast cancer cell line, SKBR3, while IgY activity was confirmed by ELISA assay using the VP2 antigen. Competition assay with native Herceptin antibodies demonstrated that the binding availability of the multimer formulation remained unaffected. Under long incubation periods, IgG multimers retained five times more activity than native IgG. In conclusion, the multimeric antibody formulations can serve as a storage depositories and sustained-release particles. These two important characteristics make this formulation promising for future novel administration protocols and altogether bring to light a different conceptual approach for the future use of therapeutic proteins as self-delivery entities rather than conjugated/encapsulated to other bio-compounds.
a Shamir Research Institute, University of Haifa , Kazrin , Israel.|b Department of Clinical Laboratory , Zefat Academic College , Zefat , Israel.|c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.
Yaron
Y
Dekel
d Consultant, specialist in the fields of biochemistry, molecular biology and genetics.
Yossy
Y
Machluf
c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.|e MIGAL - Galilee Technology Center , Kiryat Shmona , Israel.
Tal
T
Gefen
f Department of Biochemistry and Molecular Biology, Tel Aviv University, Tel Aviv, Israel , and.
Gennady
G
Eidelshtein
f Department of Biochemistry and Molecular Biology, Tel Aviv University, Tel Aviv, Israel , and.
Alexander
A
Kotlyar
g Department of Molecular Microbiology and Biotechnology , Tel Aviv University , Tel Aviv , Israel.
Yaron
Y
Bram
c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.|e MIGAL - Galilee Technology Center , Kiryat Shmona , Israel.
Ehud
E
Shahar
c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.|e MIGAL - Galilee Technology Center , Kiryat Shmona , Israel.
Farah
F
Reslane
c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.|e MIGAL - Galilee Technology Center , Kiryat Shmona , Israel.
Elina
E
Aizenshtein
c Department of Life Sciences , Tel Hai College , Upper Galilee , Israel.|e MIGAL - Galilee Technology Center , Kiryat Shmona , Israel.
Jacob
J
Pitcovski
D000906:Antibodies / Q000008:administration & dosage* / Q000737:chemistry*; D001943:Breast Neoplasms / Q000188:drug therapy*; D045744:Cell Line, Tumor; D002626:Chemistry, Pharmaceutical / Q000379:methods; D003692:Delayed-Action Preparations / Q000008:administration & dosage / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007136:Immunoglobulins / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D000068878:Trastuzumab / Q000008:administration & dosage* / Q000737:chemistry*
D000906:Antibodies; D003692:Delayed-Action Preparations; D007074:Immunoglobulin G; D007136:Immunoglobulins; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C016512:IgY; C508053:ERBB2 protein, human
Protein drug release; antibodies; bioactivity; multimers; self-delivery