Preparation and composition of inter-alpha inhibitor proteins from blood
Inventors
Lim, Yow-Pin • Sirya, Edward S. • Brne, Peter
Assignees
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Publication Number
US-9758570-B2
Publication Date
2017-09-12
Expiration Date
Abstract
The present invention generally provides processes for purification of Inter-alpha inhibitor proteins (IαIp) and compositions thereof from blood.
Core Innovation
The invention relates to purified inter-alpha inhibitor proteins (IαIps) obtained from blood/plasma and characterized by increased binding to an anti-IαIp antibody in a competitive Enzyme-Linked Immunosorbent Assay (ELISA). In particular, the IαIps are compared to IαIps not exposed to a wash buffer having a pH of 4.0 or lower, and the purified IαIps show increased competitive ELISA binding to anti-IαIp monoclonal antibodies, including MAb 69.26 and MAb 69.31.
The purification uses chromatography, including a low-pH wash at pH ≤ 4.0, with exemplified low-pH values including 3.3 and 2.9. Optionally, a high-salt wash is also used, with high-salt values exemplified at ≥250 mM NaCl, such as 290 mM. The resulting IαIps are purified to an apparent molecular weight range of about 60 to about 280 kDa and are described as having high purity and yield, exemplified as 85% to 100%.
The purified IαIps maintain trypsin inhibitory biological activity after low-pH exposure, as assessed using a trypsin inhibitory assay format. The disclosed compositions are suitable for administration to a human and are provided as pharmaceutical compositions, including kit concepts for therapeutic, prophylactic, and analytical use.
Claims Coverage
The provided material discloses one independent claim. The inventive features focus on a human-suitable composition and on characterization of IαIps by increased anti-IαIp binding in a competitive ELISA after low-pH exposure (pH 4.0 or lower), with dependent-claim refinements to composition ranges and related characteristics.
Human-suitable high-purity IαIp composition
A composition comprising 85% to 100% inter-alpha inhibitor proteins (IαIps) by weight, suitable for administration to a human.
Competitive ELISA binding increase after pH ≤4.0 exposure
IαIps characterized as having increased binding to an anti-IαIp antibody selected from MAb 69.26 and MAb 69.31 in a competitive Enzyme-Linked Immunosorbent Assay (ELISA) compared to IαIps not exposed to a wash buffer having a pH of 4.0 or lower.
The core inventive concept is a human-suitable composition containing 85% to 100% IαIps by weight, where the IαIps show increased competitive ELISA binding to anti-IαIp antibodies MAb 69.26 or MAb 69.31 relative to IαIps not exposed to a wash buffer at pH ≤4.0.
Stated Advantages
Increased binding to an anti-IαIp antibody selected from MAb 69.26 and MAb 69.31 in a competitive ELISA compared to IαIps not exposed to a wash buffer having pH 4.0 or lower.
Purified IαIps maintain trypsin inhibitory biological activity after low-pH exposure.
High purity and yield of IαIps, exemplified as 85% to 100%.
Documented Applications
Therapeutic and prophylactic use via pharmaceutical compositions suitable for administration to a human.
Analytical use via kit concepts.
Treatment indications stated in the document include sepsis/systemic inflammatory response syndrome, severe shock/septic shock, acute inflammatory diseases including rheumatoid arthritis, cancer/metastasis, infectious disease, and preterm labor.
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