title: Separation of haemocyte subpopulations in shrimp Fenneropenaeus chinensis by immunomagnetic bead using monoclonal antibody against granulocytes.
abstract: In our previous work, two monoclonal antibodies (Mabs) against granulocytes of shrimp (Fenneropenaeus chinensis) had been produced, in this paper, haemocyte subpopulations were analyzed by flow cytometry (FCM) using the Mabs. Then immunomagnetic bead (IMB) method was applied for separation hyalinocytes and granulocytes using the Mabs. The separated hyalinocytes and granulocytes were analyzed by FCM, indirect immunofluorescence assay, Giemsa staining and transmission electron microscopy, respectively. The results showed the proportion of hyalinocytes in haemolymph of F. chinensis was 15.14 ± 1.22%, and that of granulocytes was 75.43 ± 2.31%. After two times separation by IMB, the purity rate of hyalinocytes and granulocytes was 96.27 ± 1.06% and 98.13 ± 0.86%, respectively. The hyalinocytes possessed 0.60-0.85 in nucleus/cytoplasm (N/C) ratio and had few granule in cytoplasm, whereas the separated granulocytes with N/C ratio of 0.12-0.36 and high electronic density of double membrane granules. The results reported the separation of haemocyte subpopulations using Mabs in shrimp for the first time, and the hyalinocytes and granulocytes isolated by IMB could be used for their differential protein analysis.
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xing
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Yanhong
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Tang
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Xiuzhen
authors.initials: X
authors.lastname: Sheng
authors.affiliation: Laboratory of Pathology and Immunology of Aquatic Animals, KLM, Ocean University of China, Qingdao 266003, PR China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, No. 1 Wenhai Road, Aoshanwei Town, Jimo, Qingdao, PR China. Electronic address: wbzhan@ouc.edu.cn.
authors.forename: Wenbin
authors.initials: W
authors.lastname: Zhan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood; D006098:Granulocytes / Q000166:cytology / Q000276:immunology; D006434:Hemocytes / Q000166:cytology / Q000276:immunology; D018189:Immunomagnetic Separation; D033561:Penaeidae / Q000166:cytology* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal
keywords: Fenneropenaeus chinensis; Granulocytes; Hyalinocytes; Immunomagnetic bead; Monoclonal antibody