以膜鑲嵌E 型免疫球蛋白上CεmX 區域作為治療標的之實用性探究

簡易檢索 / 詳目顯示

回結果列表

研究生：	吳哲豪 Wu, Che-Hao
論文名稱：	以膜鑲嵌E 型免疫球蛋白上CεmX 區域作為治療標的之實用性探究 Studies to Investigate the Feasibility of the CεmX Domain of Membrane-bound IgE as A Therapeutic Target
指導教授：	張子文 Chang, Tse-Wen 徐邦達 Hsu, Ban-Da
口試委員:	羅傅倫 Roffler, Steve R. 張子文徐邦達莊樹諄楊立威
學位類別：	博士 Doctor
系所名稱：	生命科學暨醫學院 - 生物資訊與結構生物研究所 Institute of Bioinformatics and Structural Biology
論文出版年：	2011
畢業學年度：	100
語文別：	英文
論文頁數：	144
中文關鍵詞：	膜鑲嵌E 型免疫球蛋白、靈長類、基因發生學、單株抗體、抗CεmX 、人源化
外文關鍵詞：	CεmX, Primates, Pylogenetics, Monoclonal antibody, anti-CεmX, Humanization
相關次數：	點閱：3 下載：0
分享至:	分享至facebook 分享至twitter

查詢本校圖書館目錄查詢臺灣博碩士論文知識加值系統勘誤回報

論文內容可分成兩部分：一為探討CεmX蛋白區域之基因發生。旨在希望藉此瞭解CεmX在調節E型免疫球蛋白的角色，替抗CεmX抗體找出適合的靈長動物試驗模型。CεmX長52個胺基酸，僅表現於B淋巴細胞表面，位在膜鑲嵌E型免疫球蛋白重鏈恆常第四區域與膜錨著片段之間。膜鑲嵌免疫球蛋白對B淋巴細胞的生存與分化不可或缺。經由訊息核糖核酸的替代性剪接，依膜鑲嵌E型免疫球蛋白有或無CεmX分為長或短型，長型在試管及人體內的表現皆百倍以上顯著優於短型。CεmX擁有獨特基因和胜肽序列，已知資料庫中均無法找到類似匹配。根據小鼠和先前零星靈長動物E型免疫球蛋白重鏈基因定序結果，我們推論CεmX可能僅見於靈長動物而不存在於其他哺乳動物。小鼠及人類其他各型免疫球蛋白重鏈基因裡沒有相對應的片段，比較兩物種間各型免疫球蛋白重鏈基因膜錨著片段則發現E型免疫球蛋白的序列相似度最低，僅62.7%，明顯低於M/G/A型的92.2/88.5/77.2%。文獻顯示小鼠和大鼠在與人類CεmX相對位置的插入子基因片段序列具和相鄰表現子(重鏈第四區域與膜錨著片段)相近的類似度。綜合以上線索，我們以靈長類演化時程為經，E型免疫球蛋白重鏈基因序列為緯，展開基因發生學研究追溯CεmX和膜錨著片段的演化蹤跡。分析共26種靈長目動物及12種、7個非靈長目哺乳綱動物後，得出以下結論：1)現存三大系群趺猴的E型免疫球蛋白基因序列中，大出意外地無法標定出有意義的膜錨著片段；2)早期演化的簡鼻靈長動物，例如指猴、狐猴、懶猴的膜鑲嵌E型免疫球蛋白基因不具有CεmX，其基因型態與齧齒目等非靈長哺乳動物中所見相同；3)晚期出現的平鼻靈長動物中，新世界猴的基因序列足使人相信牠們體內只產生長膜鑲嵌E型免疫球蛋白，而不似舊世界猴、猿和人類可產生長或短型兩種。
在論文第二部份呈現將人鼠嵌合型抗CεmX單株抗體4B12人源化的步驟和結果。CεmX序列和表現細胞的獨特性使它成為以免疫學方法治療過敏疾病的優良標的，經抗體效能測試，嵌合型4B12單株抗體能成功引致表現人類長膜鑲嵌E型免疫球蛋白可結晶區的Ramos轉染B淋巴細胞株的細胞凋亡和抗體細胞毒殺作用，且使體外培養的人類週邊血B淋巴細胞E型免疫球蛋白產量下降。以(抗原)配合決定區移植法進行單株抗體人源化的技巧關鍵在選擇與小鼠源抗體基因變異部最契合的人源抗體基因變異部模版，先依序重組小鼠源抗體變異部內之配合決定區和人源框架區基因，再循理性地設計並製備候選抗體群，包含一系列人源框架區位址的胺基酸改回小鼠源框架區胺基酸的返突變。繼免疫酵素分析法相競各株候選抗體之抗原親和力於嵌合型4B12，結果顯示小鼠源抗體輕鏈變異部配合決定區移植入人源模版基因不影響抗體活性；相對之重鏈變異部移植則在第71號位址，由人源纈胺酸返突變為小鼠源精胺酸所獲得的抗體親和力最佳。Biacore生物感應器測定人源化4B12抗體親和力常數達0.98±0.21 nM略高於嵌合型的1.45±0.17 nM。

There are two parts in my dissertation. In the first part, we conducted a phylogenetic study of CεmX segment in primates to elucidate its evolutionary origin. It is hoped that this study will help to understand the function of CεmX in the regulation of IgE production. The study may also help to identify appropriate primate species as animal models to investigate anti-CεmX antibodies as potential therapeutics for IgE-mediated allergic diseases. Membrane-bound IgE (mIgE) on B lymphocytes is essential for IgE production. Earlier studies showed that the ε chain of mIgE (mε) on human B cells has a “long” isoform, with an
extra “CεmX” domain of 52 amino acid residues between the CH4 domain and the membrane anchor segment, as compared to the conventional “short” isoform. CεmX is found only in humans and a few primate species that we had examined (unpublished data), not in other species, and its sequence shares no significant homology with sequences in contemporary DNA and protein databases. Comparing membrane exons of each Ig isotype between mouse and human, the mε is the least conservative. Interestingly, 3’ region of gene segments between the CH4 and M1, correspond to the 156bp segment encoding CεmX, have high degrees of sequence
conservation in mouse and human, which is with comparable degrees to neighboring coding regions. We analyzed the ε Ig gene; in particular, its membrane exon regions encoding the membrane anchor peptide segment and CεmX domain, of 26 species of the order Primates and 12 species of seven non-Primate orders. Our analyses reveal the unexpected finding that the genes of three extant tarsier species do not contain the membrane exons for mIgE. Another striking finding is that early evolved Strepsirhini primates such as lemurs and lorises do not have gene segments for the long isoform, whereas New World monkeys such as marmosets and squirrel monkeys allow the transcription of only the long isoform. In Old World monkeys and apes, including humans, the ε gene allows the transcription of both isoforms.
In the second part of the thesis, I carried out the humanization of an anti-CεmX antibody, 4B12, which has been developed by a research team of which I am a member. After a series of characterizations, 4B12 has been determined as a suitable candidate for clinical development. Academia Sinica has also licensed the technology on 4B12 to a biotech firm based in Taiwan.
6 CεmX provides an antigenic site for targeting IgE expressing B cells. Functional studies have revealed that chimeric anti-CεmX monoclonal antibody (mAb), 4B12, can potentially control IgE production. I humanized it by first grafting its six complementarity-determining regions (CDRs) into human frameworks (FWs). Those residues in the FWs of VH and VL of the parental murine 4B12 that are essential for Fv dimer interaction and CDR conformation were identified by analyzing the binding affinity of a number of designed antibody variants, using competitive ELISA and SPR (surface plasmon resonance) analysis. A humanized 4B12
retaining the affinity of its parent was obtained by adopting all FWs of the selected human Vκ template and all FWs of the selected VH with only one residue change at position 71. The Val of the human VH was replaced by Arg at the corresponding position of the murine VH. The KD of the chimeric and humanized 4B12 are 0.98±0.21 nM and 1.45±0.17 nM, respectively.

誌謝辭    1
中文摘要    3
英文摘要    5
Chapter 1    Phylogenetic Study of Membrane-Bound IgE in Primates    7
1.1    Rationale and Significance    7
1.2    Materials and Methods    9
PCR and DNA sequencing of Primate Ig heavy ε chain (IGHE) genes    9
Analyzing IGHE loci by nucleotide BLAST    9
Isolating ε gene-harboring clones in tarsier λ phage genomic libraries by plaque hybridization screening    9
1.3    Results    11
Strepsirhini primates and non-primate mammals do not have a gene segment for the long mε isoform    11
New World monkeys, Old World monkeys and apes possess me.p    12
New World monkeys may not have the short mε isoform    12
Old World monkeys express predominantly long mε isoform    12
Tarsiers do not seem to have membrane exons of the ε Ig gene    13
1.4    Discussion    15
Chapter 2    Humanization of anti-CεmX monoclonal antibody 4B12    18
2.1    Introduction    18
2.2    Materials and Methods    21
Expression of chimeric and humanized 4B12 mAbs    21
Conjugation of chimeric 4B12 to Horseradish Peroxidase (HRP)    21
Evaluation of humanized 4B12 mAb variants by competitive ELISA    21
Computer molecular modeling of murine 4B12 Fv fragment    22
Surface plasmon resonance assay    22
2.3    Results    23
2.4    Discussion    25
Tables    26
Table I    GenBank entries and sequences of IGHE gene specific primer    26
Table II    Sources of the DNA samples analyzed in this study and the primers employed to amplify and clone the gene segments of interest..    29
Table III    Alignment programs and parameter sets employed in preparing Figure 3    32
Table IV    Species and □ gene segments analyzed in this dissertation    33
Table V    Occurrence of mε isoforms in species analyzed in this study    36
Table VI    Binding of 11-67 of chimeric and humanized 4B12 variants    38
Table VII    Effect of the VH V071R substitution by homology modeling    39
Table VIII    The kinetic constants of the binding of chimeric and humanized 4B12 to 11-67    40
Figures    41
Figure 1    Schematic representations of the structural domains and segments of the short and long isoforms of mε..    41
Figure 2    The phylogeny and molecular time scales of the evolution of primates studied in this dissertation.    42
Figure 3    Alignment of exons and homologous segments of the □ gene among the species studied in this paper.    43
Figure 4    (a) Amino acid sequence alignments of CεmX and an N-terminal segment of M1 of representative anthropoid species. The shaded segment represents the junction of CεmX and M1. (b) Amino acid and nucleotide sequence alignments at the junction of CεmX/me.p and M1/me.1, as shaded in Figure 4a..    44
Figure 5    (a) Restriction enzyme maps of the □ genomic segment of the three tarsier species studied in this paper and the loss of canonical splicing donor site at CH4. (b) Degrees of homology of DNA sequence among the three tarsier species.    45
Figure 6    Location and sequence of CεmX domain in mIgE.    46
Figure 7    Amino acid sequences of the VL and VH chains, its respective humanizing templates, and grafting of human FWs of murine 4B12.    47
Figure 8    The side view (a) and top view (b) of modeling diagrams of murine 4B12 Fv fragment.    48
Figure 9    Biacore sensograms of chimeric (a) and humanized (b) 4B12 binding to 11-67    49
Figure 10    Structural views of the humanized 4B12 Fab..    51
Appendix    52
Table A1    Unpublished diversities in the IgE gene of primates in this study    52
Figure A1    Comparison of the long and short forms of Cε1 genes in patas monkeys (E. patas) where the insertion of repetitive twelve basepairs “GCCTCCGATGCC” which encodes the polypeptide “ASDA”..    53
Figure A2    A shorter me.p segment found in the me.p of three-striped night monkeys.    54
Figure A3    Sequence diversity observed in the me.1 and me.2 of the red-bellied titi.    55
Figure A4    The intron between me.1 and me.2 of lar gibbons (H. lar) is a GC-AG intron.    56
Figure A5    A cytidine base deletion leads to frameshift mutation in siamangs’ me.2.    57
References    58
Publications    64

                                

Achatz G, Lamers M, Crameri R (2008) Membrane bound IgE: the key receptor to restrict high IgE levels. Open Immunology Journal 1:25-32.
Achatz G, Nitschke L, Lamers MC (1997) Effect of transmembrane and cytoplasmic domains of IgE on the IgE response. Science (New York, NY 276 (5311):409-411.
Anand S, Batista FD, Tkach T, Efremov DG, Burrone OR (1997) Multiple transcripts of the murine immunoglobulin epsilon membrane locus are generated by alternative splicing and differential usage of two polyadenylation sites. Molecular immunology 34 (2):175-183.
Aveskogh M, Hellman L (1995) A single major transcript encodes the membrane-bound form of rat immunoglobulin E. Scand J Immunol 42 (5):535-539.
Barrera P, Joosten LA, den Broeder AA, van de Putte LB, van Riel PL, van den Berg WB (2001) Effects of treatment with a fully human anti-tumour necrosis factor alpha monoclonal antibody on the local and systemic homeostasis of interleukin 1 and TNFalpha in patients with rheumatoid arthritis. Ann Rheum Dis 60 (7):660-669.
Batista FD, Anand S, Presani G, Efremov DG, Burrone OR (1996) The two membrane isoforms of human IgE assemble into functionally distinct B cell antigen receptors. The Journal of experimental medicine 184 (6):2197-2205.
Batista FD, Efremov DG, Burrone OR (1995) Characterization and expression of alternatively spliced IgE heavy chain transcripts produced by peripheral blood lymphocytes. J Immunol 154 (1):209-218.
Brightbill HD, Jeet S, Lin Z, Yan D, Zhou M, Tan M, Nguyen A, Yeh S, Delarosa D, Leong SR, Wong T, Chen Y, Ultsch M, Luis E, Ramani SR, Jackman J, Gonzalez L, Dennis MS, Chuntharapai A, DeForge L, Meng YG, Xu M, Eigenbrot C, Lee WP, Refino CJ, Balazs M, Wu LC (2010) Antibodies specific for a segment of human membrane IgE deplete IgE-producing B cells in humanized mice. The Journal of clinical investigation 120 (6):2218-2229.
Broide DH, Finkelman F, Bochner BS, Rothenberg ME (2011) Advances in mechanisms of asthma, allergy, and immunology in 2010. J Allergy Clin Immunol 127 (3):689-695.
Burrows B, Martinez FD, Halonen M, Barbee RA, Cline MG (1989) Association of asthma with serum IgE levels and skin-test reactivity to allergens. The New England journal of medicine 320 (5):271-277.
Burset M, Seledtsov IA, Solovyev VV (2001) SpliceDB: database of canonical and non-canonical mammalian splice sites. Nucleic acids research 29 (1):255-259.
Busse WW, Morgan WJ, Gergen PJ, Mitchell HE, Gern JE, Liu AH, Gruchalla RS, Kattan M, Teach SJ, Pongracic JA, Chmiel JF, Steinbach SF, Calatroni A, Togias A, Thompson KM, Szefler SJ, Sorkness CA (2011) Randomized trial of omalizumab (anti-IgE) for asthma in inner-city children. The New England journal of medicine 364 (11):1005-1015.
Capron A, Dessaint JP (1992) Immunologic aspects of schistosomiasis. Annu Rev Med 43:209-218.
Casale TB, Busse WW, Kline JN, Ballas ZK, Moss MH, Townley RG, Mokhtarani M, Seyfert-Margolis V, Asare A, Bateman K, Deniz Y (2006) Omalizumab pretreatment decreases acute reactions after rush immunotherapy for ragweed-induced seasonal allergic rhinitis. J Allergy Clin Immunol 117 (1):134-140.
Chang TW (2000) The pharmacological basis of anti-IgE therapy. Nat Biotechnol 18 (2):157-162.
Chang TW, Wu PC, Hsu CL, Hung AF (2007) Anti-IgE antibodies for the treatment of IgE-mediated allergic diseases. Advances in immunology 93:63-119.
Chen HY, Liu FT, Hou CM, Huang JS, Sharma BB, Chang TW (2002) Monoclonal antibodies against the C(epsilon)mX domain of human membrane-bound IgE and their potential use for targeting IgE-expressing B cells. International archives of allergy and immunology 128 (4):315-324.
Chen JB, Wu PC, Hung AF, Chu CY, Tsai TF, Yu HM, Chang HY, Chang TW (2010) Unique epitopes on C epsilon mX in IgE-B cell receptors are potentially applicable for targeting IgE-committed B cells. J Immunol 184 (4):1748-1756.
Chothia C, Lesk AM (1987) Canonical structures for the hypervariable regions of immunoglobulins. J Mol Biol 196 (4):901-917.
Cooper PJ, Ayre G, Martin C, Rizzo JA, Ponte EV, Cruz AA (2008) Geohelminth infections: a review of the role of IgE and assessment of potential risks of anti-IgE treatment. Allergy 63 (4):409-417.
Cruz AA, Lima F, Sarinho E, Ayre G, Martin C, Fox H, Cooper PJ (2007) Safety of anti-immunoglobulin E therapy with omalizumab in allergic patients at risk of geohelminth infection. Clin Exp Allergy 37 (2):197-207.
Essono S, Frobert Y, Grassi J, Creminon C, Boquet D (2003) A general method allowing the design of oligonucleotide primers to amplify the variable regions from immunoglobulin cDNA. J Immunol Methods 279 (1-2):251-266.
Green LL, Hardy MC, Maynard-Currie CE, Tsuda H, Louie DM, Mendez MJ, Abderrahim H, Noguchi M, Smith DH, Zeng Y, et al. (1994) Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs. Nat Genet 7 (1):13-21.
Hellman L (1993) Characterization of four novel epsilon chain mRNA and a comparative analysis of genes for immunoglobulin E in rodents and man. European journal of immunology 23 (1):159-167.
Holgate S, Casale T, Wenzel S, Bousquet J, Deniz Y, Reisner C (2005) The anti-inflammatory effects of omalizumab confirm the central role of IgE in allergic inflammation. J Allergy Clin Immunol 115 (3):459-465.
Horst A, Hunzelmann N, Arce S, Herber M, Manz RA, Radbruch A, Nischt R, Schmitz J, Assenmacher M (2002) Detection and characterization of plasma cells in peripheral blood: correlation of IgE+ plasma cell frequency with IgE serum titre. Clinical and experimental immunology 130 (3):370-378.
Jakobovits A, Amado RG, Yang X, Roskos L, Schwab G (2007) From XenoMouse technology to panitumumab, the first fully human antibody product from transgenic mice. Nat Biotechnol 25 (10):1134-1143.
Jones PT, Dear PH, Foote J, Neuberger MS, Winter G (1986) Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature 321 (6069):522-525.
Kaisho T, Schwenk F, Rajewsky K (1997) The roles of gamma 1 heavy chain membrane expression and cytoplasmic tail in IgG1 responses. Science (New York, NY 276 (5311):412-415.
Kaplan AP, Joseph K, Maykut RJ, Geba GP, Zeldin RK (2008) Treatment of chronic autoimmune urticaria with omalizumab. J Allergy Clin Immunol 122 (3):569-573.
Kawamura S, Kubotera N (2004) Ancestral loss of short wave-sensitive cone visual pigment in lorisiform prosimians, contrasting with its strict conservation in other prosimians. Journal of molecular evolution 58 (3):314-321.
Kawamura S, Ueda S (1992) Immunoglobulin CH gene family in hominoids and its evolutionary history. Genomics 13 (1):194-200.
Kettleborough CA, Saldanha J, Heath VJ, Morrison CJ, Bendig MM (1991) Humanization of a mouse monoclonal antibody by CDR-grafting: the importance of framework residues on loop conformation. Protein Eng 4 (7):773-783.
Kitamura D, Roes J, Kuhn R, Rajewsky K (1991) A B cell-deficient mouse by targeted disruption of the membrane exon of the immunoglobulin mu chain gene. Nature 350 (6317):423-426.
Kolbinger F, Saldanha J, Hardman N, Bendig MM (1993) Humanization of a mouse anti-human IgE antibody: a potential therapeutic for IgE-mediated allergies. Protein Eng 6 (8):971-980.
Leung DY, Sampson HA, Yunginger JW, Burks AW, Jr., Schneider LC, Wortel CH, Davis FM, Hyun JD, Shanahan WR, Jr. (2003) Effect of anti-IgE therapy in patients with peanut allergy. The New England journal of medicine 348 (11):986-993.
LoBuglio AF, Wheeler RH, Trang J, Haynes A, Rogers K, Harvey EB, Sun L, Ghrayeb J, Khazaeli MB (1989) Mouse/human chimeric monoclonal antibody in man: kinetics and immune response. Proceedings of the National Academy of Sciences of the United States of America 86 (11):4220-4224.
Major JG, Jr., Davis FM, Liou RS, Chang TW (1996) Structural features of the extracellular portion of membrane-anchoring peptides on membrane-bound immunoglobulins. Molecular immunology 33 (2):179-187.
Merker S, Driller C, Perwitasari-Farajallah D, Pamungkas J, Zischler H (2009) Elucidating geological and biological processes underlying the diversification of Sulawesi tarsiers. Proceedings of the National Academy of Sciences of the United States of America 106 (21):8459-8464.
Merker S, Driller C, Perwitasari-Farajallah D, Zahner R, Zischler H (2007) Isolation and characterization of 12 microsatellite loci for population studies of Sulawesi tarsiers (Tarsius spp.). Molecular Ecology Notes 7 (6):1216-1218.
Morea V, Lesk AM, Tramontano A (2000) Antibody modeling: implications for engineering and design. Methods 20 (3):267-279.
Nilsson L, Islam KB, Olafsson O, Zalcberg I, Samakovlis C, Hammarstrom L, Smith CI, Sideras P (1991) Structure of TGF-beta 1-induced human immunoglobulin C alpha 1 and C alpha 2 germ-line transcripts. International immunology 3 (11):1107-1115.
O'Connor SJ, Meng YG, Rezaie AR, Presta LG (1998) Humanization of an antibody against human protein C and calcium-dependence involving framework residues. Protein Eng 11 (4):321-328.
Pace E, Ferraro M, Bruno A, Chiappara G, Bousquet J, Gjomarkaj M (2011) Clinical benefits of 7 years of treatment with omalizumab in severe uncontrolled asthmatics. J Asthma 48 (4):387-392.
Peng C, Davis FM, Sun LK, Liou RS, Kim YW, Chang TW (1992) A new isoform of human membrane-bound IgE. J Immunol 148 (1):129-136.
Perelman P, Johnson WE, Roos C, Seuanez HN, Horvath JE, Moreira MA, Kessing B, Pontius J, Roelke M, Rumpler Y, Schneider MP, Silva A, O'Brien SJ, Pecon-Slattery J (2011) A molecular phylogeny of living primates. PLoS genetics 7 (3):e1001342.
Pinot de Moira A, Fulford AJ, Kabatereine NB, Ouma JH, Booth M, Dunne DW (2010) Analysis of complex patterns of human exposure and immunity to Schistosomiasis mansoni: the influence of age, sex, ethnicity and IgE. PLoS Negl Trop Dis 4 (9).
Presta LG, Chen H, O'Connor SJ, Chisholm V, Meng YG, Krummen L, Winkler M, Ferrara N (1997) Humanization of an anti-vascular endothelial growth factor monoclonal antibody for the therapy of solid tumors and other disorders. Cancer Res 57 (20):4593-4599.
Presta LG, Lahr SJ, Shields RL, Porter JP, Gorman CM, Fendly BM, Jardieu PM (1993) Humanization of an antibody directed against IgE. J Immunol 151 (5):2623-2632.
Queen C, Schneider WP, Selick HE, Payne PW, Landolfi NF, Duncan JF, Avdalovic NM, Levitt M, Junghans RP, Waldmann TA (1989) A humanized antibody that binds to the interleukin 2 receptor. Proceedings of the National Academy of Sciences of the United States of America 86 (24):10029-10033.
Reth M, Tasuku H, Frederick WA (1995) Antigen receptors on B lymphocytes. In: Immunoglobulin Genes (Second Edition). Academic Press, London, pp 129-142.
Shawler DL, Bartholomew RM, Smith LM, Dillman RO (1985) Human immune response to multiple injections of murine monoclonal IgG. J Immunol 135 (2):1530-1535.
Shekelle M, Meier R, Wahyu I, Wirdateti, Ting N (2010) Molecular Phylogenetics and Chronometrics of Tarsiidae Based on 12S mtDNA Haplotypes: Evidence for Miocene Origins of Crown Tarsiers and Numerous Species within the Sulawesian Clade. International Journal of Primatology 31 (6):1083-1106.
Shin HD, Kim LH, Park BL, Jung JH, Kim JY, Chung IY, Kim JS, Lee JH, Chung SH, Kim YH, Park HS, Choi JH, Lee YM, Park SW, Choi BW, Hong SJ, Park CS (2003) Association of Eotaxin gene family with asthma and serum total IgE. Hum Mol Genet 12 (11):1279-1285.
Stone KD, Prussin C, Metcalfe DD (2010) IgE, mast cells, basophils, and eosinophils. J Allergy Clin Immunol 125 (2 Suppl 2):S73-80.
Sun X, Hia HC, Goh PE, Yap MG (2008) High-density transient gene expression in suspension-adapted 293 EBNA1 cells. Biotechnol Bioeng 99 (1):108-116.
Tashita H, Fukao T, Kaneko H, Teramoto T, Inoue R, Kasahara K, Kondo N (1998) Molecular basis of selective IgG2 deficiency. The mutated membrane-bound form of gamma2 heavy chain caused complete IGG2 deficiency in two Japanese siblings. The Journal of clinical investigation 101 (3):677-681.
Tramontano A, Chothia C, Lesk AM (1990) Framework residue 71 is a major determinant of the position and conformation of the second hypervariable region in the VH domains of immunoglobulins. J Mol Biol 215 (1):175-182.
Vernersson M, Aveskogh M, Hellman L (2004) Cloning of IgE from the echidna (Tachyglossus aculeatus) and a comparative analysis of epsilon chains from all three extant mammalian lineages. Dev Comp Immunol 28 (1):61-75.
Vernersson M, Aveskogh M, Munday B, Hellman L (2002) Evidence for an early appearance of modern post-switch immunoglobulin isotypes in mammalian evolution (II); cloning of IgE, IgG1 and IgG2 from a monotreme, the duck-billed platypus, Ornithorhynchus anatinus. European journal of immunology 32 (8):2145-2155.
Werther WA, Gonzalez TN, O'Connor SJ, McCabe S, Chan B, Hotaling T, Champe M, Fox JA, Jardieu PM, Berman PW, Presta LG (1996) Humanization of an anti-lymphocyte function-associated antigen (LFA)-1 monoclonal antibody and reengineering of the humanized antibody for binding to rhesus LFA-1. J Immunol 157 (11):4986-4995.
Willuda J, Honegger A, Waibel R, Schubiger PA, Stahel R, Zangemeister-Wittke U, Pluckthun A (1999) High thermal stability is essential for tumor targeting of antibody fragments: engineering of a humanized anti-epithelial glycoprotein-2 (epithelial cell adhesion molecule) single-chain Fv fragment. Cancer Res 59 (22):5758-5767.
Wisdom GB (2005) Conjugation of Antibodies to Horseradish Peroxidase
Immunochemical Protocols. In: Burns R (ed), vol 295. Methods in Molecular Biology. Humana Press, pp 127-130.
Wu TT, Kabat EA (1970) An analysis of the sequences of the variable regions of Bence Jones proteins and myeloma light chains and their implications for antibody complementarity. The Journal of experimental medicine 132 (2):211-250.
Yel L, Minegishi Y, Coustan-Smith E, Buckley RH, Trubel H, Pachman LM, Kitchingman GR, Campana D, Rohrer J, Conley ME (1996) Mutations in the mu heavy-chain gene in patients with agammaglobulinemia. The New England journal of medicine 335 (20):1486-1493.
Zhang K, Saxon A, Max EE (1992) Two unusual forms of human immunoglobulin E encoded by alternative RNA splicing of epsilon heavy chain membrane exons. The Journal of experimental medicine 176 (1):233-243.

全文公開日期本全文未授權公開 (校內網路)
全文公開日期本全文未授權公開 (校外網路)

簡易檢索 / 詳目顯示

相關論文