Crude Heparin Preparations Unveil the Presence of Structurally Diverse Oversulfated Contaminants
Aline Mendes,
Maria C. Z. Meneghetti,
Marcelly Valle Palladino,
Giselle Zenker Justo,
Guilherme L. Sassaki,
Jawed Fareed,
Marcelo A. Lima,
Helena B. Nader
Affiliations
Aline Mendes
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Maria C. Z. Meneghetti
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Marcelly Valle Palladino
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Giselle Zenker Justo
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Guilherme L. Sassaki
Departamento de Bioquímica, Setor de Ciências Biológicas, Centro Politécnico, Universidade Federal do Paraná, Avenida Coronel Francisco H. dos Santos, 100, Caixa Postal 19031, Curitiba, PR 81531-980, Brazil
Jawed Fareed
Department of Molecular Pharmacology & Therapeutics, Hemostasis & Thrombosis Research Laboratories, Loyola University Medical Center, 2160 S. First Avenue Maywood, IL 60153, USA
Marcelo A. Lima
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Helena B. Nader
Departamento de Bioquímica, Instituto de Farmacologia e Biologia Molecular, Escola Paulista de Medicina, Universidade Federal de São Paulo, Rua Três de Maio, 100 – São Paulo, SP 04044-020, Brazil
Nowadays, pharmaceutical heparin is purified from porcine and bovine intestinal mucosa. In the past decade there has been an ongoing concern about the safety of heparin, since in 2008, adverse effects associated with the presence of an oversulfated chondroitin sulfate (OSCS) were observed in preparations of pharmaceutical porcine heparin, which led to the death of patients, causing a global public health crisis. However, it has not been clarified whether OSCS has been added to the purified heparin preparation, or whether it has already been introduced during the production of the raw heparin. Using a combination of different analytical methods, we investigate both crude and final heparin products and we are able to demonstrate that the sulfated contaminants are intentionally introduced in the initial steps of heparin preparation. Furthermore, the results show that the oversulfated compounds are not structurally homogeneous. In addition, we show that these contaminants are able to bind to cells in using well known heparin binding sites. Together, the data highlights the importance of heparin quality control even at the initial stages of its production.
