Itory effect of PAb on tumor growth in xenograft SCID mouse models. (A) A significant difference in tumor volume (P,0.05) was observed between PAb-treated mice and other treatment groups. The mean 6 standard error of the mean of tumor growth of five mice is shown. (B) Representative picture for tumor volume different groups. (C) A significant increase in survival was observed in PAb-treated mice compared with other treatment groups (P,0.05). doi:10.1371/journal.pone.0059117.gScreening of MM by Polyclonal ImmunoglobulinFigure 6. PAb-induced tumor cells apoptosis in vivo by TUNEL assay. 25033180 (A) Sections from the tumor-bearing mice treated with NS (left panel), control IgG (middle panel), or PAb (right panel) were stained with FITC-dUTP as described in the Materials and Methods section (2006). (B) An apparent increase in the number of apoptotic cells and apoptotic index was observed within residual tumors treated with PAb compared with other treatment groups in the ARH-77 subcutaneous injection tumor models. * represents the PAb group showing significant difference compared with NS and control IgG group mice (P,0.05). doi:10.1371/journal.pone.0059117.gDiscussionThe availability of high throughput 2-DE gels and initial screening using automated procedures has made the identification of TAA in the proteome 
of various tumor cell lines and/or tissues possible. This study was based on PAb combined with proteomic analysis and aimed to screen TAAs in the proteome level to help further improve the diagnosis and immunotherapy of MM. We synthesized a PAb by immunizing rabbits with the human plasmacytoma cell line ARH-77 and identified multiple TAAs of MM, such as enolase, ADPH, 
and HSP90s, among others, using 2-DE, Western blot, and mass spectrometric techniques. To validate the MS/MS results, we selected three proteins for examination according to their positions in the Mascot score list, which lists the vital role they play in many cancers. These proteins are discussed below.a-enolase, a key MedChemExpress Vasopressin enzyme in the glycolysis pathway, is upregulated in 18 out of 24 types of cancer, as determined by bioinformatics study using gene chips and EST databases [21]. A recent proteomic analysis further revealed that overexpression of aenolase in hepatitis C virus-related hepatocellular carcinomas is associated with tumor progression 25033180 (A) Sections from the tumor-bearing mice treated with NS (left panel), control IgG (middle panel), or PAb (right panel) were stained with FITC-dUTP as described in the Materials and Methods section (2006). (B) An apparent increase in the number of apoptotic cells and apoptotic index was observed within residual tumors treated with PAb compared with other treatment groups in the ARH-77 subcutaneous injection tumor models. * represents the PAb group showing significant difference compared with NS and control IgG group mice (P,0.05). doi:10.1371/journal.pone.0059117.gDiscussionThe availability of high throughput 2-DE gels and initial screening using automated procedures has made the identification of TAA in the proteome of various tumor cell lines and/or tissues possible. This study was based on PAb combined with proteomic analysis and aimed to screen TAAs in the proteome level to help further improve the diagnosis and immunotherapy of MM. We synthesized a PAb by immunizing rabbits with the human plasmacytoma cell line ARH-77 and identified multiple TAAs of MM, such as enolase, ADPH, and HSP90s, among others, using 2-DE, Western blot, and mass spectrometric techniques. To validate the MS/MS results, we selected three proteins for examination according to their positions in the Mascot score list, which lists the vital role they play in many cancers. These proteins are discussed below.a-enolase, a key enzyme in the glycolysis pathway, is upregulated in 18 out of 24 types of cancer, as determined by bioinformatics study using gene chips and EST databases [21]. A recent proteomic analysis further revealed that overexpression of aenolase in hepatitis C virus-related hepatocellular carcinomas is associated with tumor progression 23727046 [22]. Although the mechanisms of the surface expression and orientation of a-enolase on the membrane have yet to be clearly understood, surface a-enolase is known to act as a strong plasminogen-binding receptor [23]. The binding of plasminogen to the cell surface and its consequent activation to plasmin may play crucial roles in the intravascular and pericellular fibrinolytic systems, cell invasion, tumor cell migration, and metastasis as a plasminogen-binding receptor [24]. Thus, we hypothesize that a-enolase is a diagnostic marker and therapeutic target of MM.ADPH a-EnolaseThe propensity for glycolysis is enhanced in cancer cells because of increased cell proliferation. Previous studies have indicated thatADPH, a member of the perilipin family of lipid dropletassociated proteins, hypothetically mediates milk lipid formation and secretion [25]. Previous studies have indicated that ADPHScreening of MM by Polyclonal Immunoglobulinfunctions in lipid storage droplets formation [26], fatty acid uptake [27], and milk lipid secretion [28]. In addition, ADPH is reportedly overexpressed in colorectal cancer [29], hepatocellular cancer, renal cell c.