Extraction of superoxide dismutase, catalase, and carbonic anhydrase from stroma-free red blood cell hemolysate for the preparation of the nanobiotechnological complex of polyhemoglobin–superoxide dismutase–catalase–carbonic anhydrase

Guo, Cheng; Gynn, M; Chang, Thomas Ming Swi

doi:10.3109/21691401.2015.1035479

Cited by 19 publications

(19 citation statements)

References 23 publications

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“…In this complex, not only are all three RBC functions present, they are also enhanced due to the increased concentrations of RBC enzymes in the complex. These RBC enzymes can be extracted from RBCs inexpensively (Guo et al 2015). This complex does not contain into any blood groups.…”

mentioning

confidence: 99%

Red blood cell replacement, or nanobiotherapeutics with enhanced red blood cell functions?

Chang

2015

Artificial Cells, Nanomedicine, and Biotechnology

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mentioning

confidence: 99%

Red blood cell replacement, or nanobiotherapeutics with enhanced red blood cell functions?

Chang

2015

Artificial Cells, Nanomedicine, and Biotechnology

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“…Ongoing study also show that Poly-[SFHb-SOD-CAT-CA] prepared with the new enzyme extraction method (Guo et al 2015) also has increased temperature stability.…”

Section: Effects Of Lyophilization On Enzyme Activitymentioning

confidence: 75%

Temperature stability of Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] in the form of a solution or in the lyophilized form during storage at −80 °C, 4 °C, 25 °C and 37 °C or pasteurization at 70 °C

Bian

Guo

Chang

2015

Artificial Cells, Nanomedicine, and Biotechnology

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Polyhemoglobin-superoxide dismutase-catalase-carbonic anhydrase (Poly-[Hb-SOD-CAT-CA]) contains all three major functions of red blood cells (RBCs) at an enhanced level. It transports oxygen, removes oxygen radicals and transports carbon dioxide. Our previous studies in a 90-min 30 mm Hg Mean Arterial Pressure (MAP) sustained hemorrhagic shock rat model shows that it is more effective than blood in the lowering of elevated intracellular pCO 2 , recovery of ST-elevation and histology of the heart and intestine. This paper is to analyze the storage and temperature stability. Allowable storage time for RBC is about 1 d at room temperature and 42 d at 4C. Also, RBC cannot be pasteurized to remove infective agents like HIV and Ebola. PolyHb can be heat sterilized and can be stored for 1 year even at room temperature. However, Poly-[Hb-SOD-CAT-CA] contains both Hb and enzymes and enzymes are particularly sensitive to storage and heat. We thus carried out studies to analyze its storage stability at different temperatures and heat pasteurization stability. Results of storage stability show that lyophilization extends the storage time to 1 year at 4 C and 40 d at room temperature (compared to respectively, 42 d and 1 d for RBC). After the freeze-dry process, the enzyme activities of Poly-[SFHb-SOD-CAT-CA] was 100 ± 2% for CA, 100 ± 2% for SOD and 93 ± 3.5% for CAT. After heat pasteurization at 70 C for 2 h, lyophilized Poly-[Hb-SOD-CAT-CA] retained good enzyme activities of CA 97 ± 4%, SOD 100 ± 2.5% and CAT 63.8 ± 4%. More CAT can be added during the crosslinking process to maintain the same enzyme ratio after heat pasteurization. Heat pasteurization is possible only for the lyophilized form of Poly-[Hb-SOD-CAT-CA] and not for the solution. It can be easily reconstituted by dissolving in suitable solutions that continues to have good storage stability though less than that for the lyophilized form. According to the P50 value, Poly-[SFHb-SOD-CAT-CA] retains its oxygen carrying ability before and after long-term storage.ARTICLE HISTORY

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“…SFHb contains haemoglobin and red blood cell enzymes including CAT, SOD and CA. The enzyme extracted had been established previously [24,25]. 300 ml bovine SFHb (Hb concentration: 9g/dL) is placed in a beaker on ice.…”

Section: Bovine Enzyme Extraction and MIX With Bovine Stroma-free Haementioning

confidence: 99%

“…To enhance the red blood cell enzymes, commercial CAT, SOD and CA are added for the preparation of poly-[Hb-CAT-SOD-CA] [20][21][22][23]. For avoiding the high cost of commercial enzymes, a cost-effective novel enzyme extraction method has been established to obtain these three enzymes from SFHb simultaneously [24,25]. This avoids the need for expensive commercial enzymes and allows the large-scale production of poly-[Hb-CAT-SOD-CA] in the future.…”

Section: Introductionmentioning

confidence: 99%

Long term safety and immunological effects of a nanobiotherapeutic, bovine poly-[hemoglobin-catalase-superoxide dismutase-carbonic anhydrase], after four weekly 5% blood volume top-loading followed by a challenge of 30% exchange transfusion

Guo

Chang

2018

Artificial Cells, Nanomedicine, and Biotechnology

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The long-term safety and immunological effects of bovine poly-[haemoglobin-catalase-superoxide dismutase-carbonic anhydrase] in rats are studied by four-weekly 5% blood volume top-loading infusions followed by 30% blood volume exchange transfusion. There is no significant difference in growth, biochemistry and blood pressure between the control group receiving lactated ringer solution and those receiving the bovine poly-[haemoglobin-catalase-superoxide dismutase-carbonic anhydrase]. There is no significant change in mean arterial pressures (MAP) before and after each weekly top-loading infusion. After both the four weekly top-loading and the 30% exchange transfusions, the following safety and immune response evaluations are carried out. These include general studies on Ouchterlony double diffusion, total IgG and IgM, and complement activation. This is followed by quantitative measurements of specific antibodies against each of the following bovine components: Hb, CAT, SOD and CA in bovine poly-[haemoglobin-catalase-superoxide dismutase-carbonic anhydrase]. After the four weekly top-loading, each rat received a challenge of 30% blood volume exchange transfusion. The MAP, histamine and tryptase levels are tested before and after the 30% exchange transfusion. There are no anaphylactic reactions as shown by the MAP or histamine and tryptase. The results showed no safety problem nor adverse immune responses. All the rats survived when followed for one week after the 30% exchange transfusion.

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Extraction of superoxide dismutase, catalase, and carbonic anhydrase from stroma-free red blood cell hemolysate for the preparation of the nanobiotechnological complex of polyhemoglobin–superoxide dismutase–catalase–carbonic anhydrase

Cited by 19 publications

References 23 publications

Red blood cell replacement, or nanobiotherapeutics with enhanced red blood cell functions?

Red blood cell replacement, or nanobiotherapeutics with enhanced red blood cell functions?

Temperature stability of Poly-[hemoglobin-superoxide dismutase-catalase-carbonic anhydrase] in the form of a solution or in the lyophilized form during storage at −80 °C, 4 °C, 25 °C and 37 °C or pasteurization at 70 °C

Long term safety and immunological effects of a nanobiotherapeutic, bovine poly-[hemoglobin-catalase-superoxide dismutase-carbonic anhydrase], after four weekly 5% blood volume top-loading followed by a challenge of 30% exchange transfusion

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