Last updated: 2020-06-24

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Data input

Read and annotate data

Process raw data from LUMOS

rawTab <- read_tsv("../data/LUMOS/20191113_112707_20191016_p3254_o6152_hybrid_lybrary_all_Report_proteinQuant.tsv") %>% 
  separate(R.FileName, c("a","b","c","d","id","e"), sep = "_") %>%
  select(id, PG.ProteinAccessions, PG.Quantity) %>% 
  mutate(id = paste0("A_1_",id)) %>%
  dplyr::rename(ID = PG.ProteinAccessions, 
                count = PG.Quantity)

#annotate patient ID
patAnno <- readxl::read_xlsx("../data/SampleAnnotation_cleaned.xlsx") %>%
  mutate(id = paste0("A_1_",id)) %>%
  select(-Institute, -Source, -diagnosis)

#annotate basic genomic feature
genAnno <- patMeta %>% select(Patient.ID, gender, IGHV.status, trisomy12) %>%
  mutate(trisomy12 = as.integer(as.character(trisomy12))) %>%
  mutate(trisomy12 = factor(trisomy12))

#annotate technical variable
techTab <- readxl::read_xlsx("../data/20191025_Proteom_submitted_samples_final.xlsx") %>%
  select(`Patient ID`, operator, viability, batch, `date of sample processing`, `protein conc. in ug`, `freeze-thaw cycles of peptide solution`) %>% dplyr::rename(patID = `Patient ID`, processDate = `date of sample processing`, proteinConc = `protein conc. in ug`, `freeThawCycle` = `freeze-thaw cycles of peptide solution`) %>%
  mutate(batch = ifelse(batch == "test run", "0", batch))

patAnno <- left_join(patAnno, genAnno, by = c(patID = "Patient.ID")) %>%
  left_join(techTab, by = "patID")

#update protein concentration using new data
protConc <- readxl::read_xlsx("../data/LUMOS/p3254_o6152_Nanodrop_measurement.xlsx") %>% mutate(id = paste0("A_1_", sample)) %>%
  mutate(patID = patAnno[match(id, patAnno$id),]$patID)
patAnno$proteinConc <- protConc[match(patAnno$patID, protConc$patID),]$`Nanodrop measurement`


rawTab <- left_join(rawTab, patAnno, by = "id") %>% dplyr::rename(name=ID)

Generate formated protein ID

idMap <- tibble(name = unique(rawTab$name)) %>% 
  mutate(ID = paste0("prot",seq(nrow(.))))
rawTab <- left_join(rawTab, idMap, by = "name")

Create SummarizedExperiment object

protCLL <- tidyToSum(rawTab, "ID", "patID", "count", 
                     annoCol = colnames(patAnno)[colnames(patAnno) != "patID"],
                     annoRow = c("name"))
rowData(protCLL)$ID <- rownames(protCLL)

Dimension of the original data

dim(protCLL)
[1] 4452   50

Initial QA

Check distribution of missing values

Pattern of missing values

plot_missval(protCLL)

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Some samples show less detection rate than others.

Frequencies of missing values for each sample

proTab <- sumToTiday(protCLL, "uniprotID", "patientID")
patMiss <- group_by(proTab, patientID) %>%
  summarise(freqNA = sum(is.na(count))/length(count)) %>%
  arrange(desc(freqNA)) %>% 
  mutate(patientID = factor(patientID, levels = patientID))

ggplot(patMiss, aes(x = patientID, y = freqNA)) + geom_point(size=3) + 
  geom_segment(aes(x=patientID, xend=patientID, y=0, yend=freqNA)) + 
  theme(axis.text.x = element_text(angle = 90, vjust =0.5, hjust=1)) + ylab("Frenquency")

Version Author Date
cc8c163 Junyan Lu 2020-02-27

One sample P0641, has much more missing values than other samples. This sample also behaves strangely in down-stream analysis.

Remove “P0641”

protCLL <- protCLL[,colnames(protCLL)!="P0641"]
proTab <- sumToTiday(protCLL, "uniprotID", "patientID")

Proteins with missing values

proMiss <- group_by(proTab, uniprotID) %>%
  summarise(freqNA = sum(is.na(count))/length(count)) %>%
  arrange(desc(freqNA)) %>% 
  mutate(uniprotID = factor(uniprotID, levels = uniprotID))

head(proMiss)
# A tibble: 6 x 2
  uniprotID freqNA
  <fct>      <dbl>
1 prot4425   0.980
2 prot4430   0.980
3 prot4433   0.980
4 prot4434   0.980
5 prot4435   0.980
6 prot4436   0.980
ggplot(proMiss, aes(x=freqNA)) + geom_histogram() +
  xlab("Missing value frequency")
`stat_bin()` using `bins = 30`. Pick better value with `binwidth`.

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Missing value cut-off versus number of remaining proteins

sumTab <- lapply(seq(0,1,by = 0.01), function(x) tibble(cut = x, freq = sum(proMiss$freqNA < x)/nrow(proMiss))) %>% bind_rows()
ggplot(sumTab, aes(x=cut, y=freq)) + geom_line() + xlab("Missing value cut-off") + ylab("Percent remaining")

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Missing value frequency versus median expression

compareTab <- group_by(proTab, uniprotID) %>%
  summarise(freqNA = sum(is.na(count))/length(count),
            medianExpr = median(log2(count), na.rm=TRUE))

ggplot(compareTab, aes(x=freqNA, y = medianExpr)) + geom_point() + geom_smooth(method = "loess") +
  ylab("Median log2 count") + xlab("Frequency of missing values")

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Highly expressed proteins tend to have higher detection rate.

Remove proteins with more than 50% missing values

cut=0.5
protCLL_filt <- protCLL[rowSums(is.na(assay(protCLL)))/ncol(protCLL) <= cut,]

Dimension of the filtered data

dim(protCLL_filt)
[1] 4291   49

Data normalization

Distribution of raw data

protTab <- sumToTiday(protCLL_filt, "id","patientID")
ggplot(protTab, aes(x=patientID, y=count)) + geom_boxplot() + scale_y_log10() +
  theme(axis.text.x = element_text(angle = 90, vjust = 0.5, hjust=1))
Warning: Removed 5902 rows containing non-finite values (stat_boxplot).

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Normalize data using vsn package

exprMat <- assay(protCLL_filt)
resVsn <- vsnMatrix(exprMat)
protCLL_norm <- protCLL_filt
assay(protCLL_norm) <- predict(resVsn, exprMat)

Boxplot of normalized counts

protTab <- sumToTiday(protCLL_norm, "uniprotID","patientID")
ggplot(protTab, aes(x=patientID, y=count)) + geom_boxplot() +
  theme(axis.text.x = element_text(angle = 90, vjust = 0.5, hjust=1))
Warning: Removed 5902 rows containing non-finite values (stat_boxplot).

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Mean VS SD plot after normalization

vsn::meanSdPlot(resVsn)

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Looks OK. Although lowly expressed proteins still have higher variance.

Post processing

Impute missing values

For impute missing values, first I need to see whether the data is missing at random or not.

Check missing value pattern again (random or not?)

Missing value pattern after normalization and filtering

plot_missval(protCLL_norm)

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Detection rate of proteins with and without missing values

plot_detect(protCLL_norm)

Version Author Date
cc8c163 Junyan Lu 2020-02-27

Proteins with missing values have on average low intensities. Not missing at random.

Impute missing values

Impute missing values using quantile regression-based left-censored function (QRILC)

This is a method for imputing missing not at random data.

protCLL_imp <- impute(protCLL_norm, fun = "QRILC")
#add QRILC imputed data
assays(protCLL_norm)[["QRILC"]] <- assay(protCLL_imp)

Annotate protein information using bioMart

Prepare protein id table

idTab <-   map_df(unique(rowData(protCLL)$name), ~tibble(ID = ., uniprotID = str_split(.,";")[[1]]))
ensembl = useMart("ensembl",dataset="hsapiens_gene_ensembl",host="grch37.ensembl.org")

ids <- idTab$uniprotID
anno <- getBM(attributes=c('hgnc_symbol','ensembl_gene_id',
                           'uniprotswissprot'), 
      filters = 'uniprotswissprot', 
      values = ids, 
      mart = ensembl)

idTab <- left_join(idTab, anno, by = c(uniprotID="uniprotswissprot"))

Proteins that can not be mapped

sort(unique(filter(idTab, is.na(ensembl_gene_id))$uniprotID))
  [1] "A0A024RBG1"      "A0A075B6I0"      "A0A075B6K4"     
  [4] "A0A075B6P5"      "A0A0A0MT36"      "A0A0A0MTA4"     
  [7] "A0A0B4J1X5"      "A0A0B4J1X8"      "A0A0B4J2A2"     
 [10] "A0A0B4J2D5"      "A0A0B4J2D9"      "A0A0C4DH29"     
 [13] "A0A0C4DH31"      "A0A0C4DH38"      "A0A0C4DH67"     
 [16] "A0A0C4DH69"      "A0A0J9YXX1"      "A6NI72"         
 [19] "A8MWD9"          "C4AMC7"          "E9PAV3"         
 [22] "E9PRG8"          "L0R6Q1"          "O15126"         
 [25] "O75396"          "P01593"          "P01594"         
 [28] "P01597"          "P01599"          "P01602"         
 [31] "P01615"          "P01619"          "P01624"         
 [34] "P01701"          "P01709"          "P01718"         
 [37] "P01742"          "P01764"          "P01780"         
 [40] "P01782"          "P01850"          "P01871"         
 [43] "P01877"          "P01880"          "P04430"         
 [46] "P04432"          "P06331"          "P0C7X5"         
 [49] "P0CB33"          "P0CF74"          "P0DMM9"         
 [52] "P0DMN0"          "P0DMV8"          "P0DMV9"         
 [55] "P0DN76"          "P0DOY2"          "P0DOY3"         
 [58] "P0DP02"          "P0DP04"          "P0DP09"         
 [61] "P0DP23"          "P0DP24"          "P0DP25"         
 [64] "P0DPI2"          "P16284"          "P21127"         
 [67] "P58107"          "P80748"          "Q58FF6"         
 [70] "Q58FG1"          "Q59GN2"          "Q6EEV4"         
 [73] "Q6SZW1"          "Q6VEQ5"          "Q6ZSR6"         
 [76] "Q6ZSR9"          "Q8IY67"          "Q8NHM4"         
 [79] "Q96M42"          "Q9BXJ8"          "Q9BYP9"         
 [82] "Q9H270"          "Q9NPI6"          "Q9Y2S7"         
 [85] "Y-FGCZCont00001" "Y-FGCZCont00010" "Y-FGCZCont00024"
 [88] "Y-FGCZCont00027" "Y-FGCZCont00030" "Y-FGCZCont00032"
 [91] "Y-FGCZCont00033" "Y-FGCZCont00035" "Y-FGCZCont00037"
 [94] "Y-FGCZCont00052" "Y-FGCZCont00054" "Y-FGCZCont00056"
 [97] "Y-FGCZCont00060" "Y-FGCZCont00064" "Y-FGCZCont00065"
[100] "Y-FGCZCont00066" "Y-FGCZCont00077" "Y-FGCZCont00093"
[103] "Y-FGCZCont00098" "Y-FGCZCont00099" "Y-FGCZCont00100"
[106] "Y-FGCZCont00106" "Y-FGCZCont00110" "Y-FGCZCont00111"
[109] "Y-FGCZCont00113" "Y-FGCZCont00116" "Y-FGCZCont00118"
[112] "Y-FGCZCont00122" "Y-FGCZCont00124" "Y-FGCZCont00127"
[115] "Y-FGCZCont00142" "Y-FGCZCont00147" "Y-FGCZCont00148"
[118] "Y-FGCZCont00150" "Y-FGCZCont00151" "Y-FGCZCont00160"
[121] "Y-FGCZCont00161" "Y-FGCZCont00162" "Y-FGCZCont00165"
[124] "Y-FGCZCont00168" "Y-FGCZCont00175" "Y-FGCZCont00180"
[127] "Y-FGCZCont00188" "Y-FGCZCont00189" "Y-FGCZCont00190"
[130] "Y-FGCZCont00193" "Y-FGCZCont00195" "Y-FGCZCont00212"
[133] "Y-FGCZCont00231" "Y-FGCZCont00237" "Y-FGCZCont00243"
[136] "Y-FGCZCont00248" "Y-FGCZCont00251" "Y-FGCZCont00256"
[139] "Y-FGCZCont00260" "Y-FGCZCont00272" "Y-FGCZCont00276"
[142] "Y-FGCZCont00278" "Y-FGCZCont00282" "Y-FGCZCont00285"
[145] "Y-FGCZCont00286" "Y-FGCZCont00288" "Y-FGCZCont00289"
[148] "Y-FGCZCont00293" "Y-FGCZCont00301" "Y-FGCZCont00307"
[151] "Y-FGCZCont00310" "Y-FGCZCont00311" "Y-FGCZCont00319"
[154] "Y-FGCZCont00328" "Y-FGCZCont00329" "Y-FGCZCont00330"
[157] "Y-FGCZCont00343" "Y-FGCZCont00347" "Y-FGCZCont00348"
[160] "Y-FGCZCont00351" "Y-FGCZCont00355" "Y-FGCZCont00356"
[163] "Y-FGCZCont00359" "Y-FGCZCont00367" "Y-FGCZCont00373"
[166] "Y-FGCZCont00374" "Y-FGCZCont00375" "Y-FGCZCont00376"
[169] "Y-FGCZCont00378" "Y-FGCZCont00380" "Y-FGCZCont00414"
[172] "Y-FGCZCont00417" "Y-FGCZCont00418" "Y-FGCZCont00419"
[175] "Y-FGCZCont00422" "Y-FGCZCont00459" "Y-FGCZCont00461"
[178] "Y-FGCZCont00463" "Y-FGCZCont00474" "Y-FGCZCont00482"

Map those proteins using uniprotID to symbol list

idTab.miss <- filter(idTab,is.na(ensembl_gene_id))
mapSymbol <- read_tsv("../data/mapSymbol.txt")
Parsed with column specification:
cols(
  From = col_character(),
  To = col_character()
)
idTab.miss <- mutate(idTab.miss, hgnc_symbol = mapSymbol[match(uniprotID, mapSymbol$From),]$To)
#proteins that can not be mapped
unique(filter(idTab.miss, is.na(hgnc_symbol))$uniprotID)
 [1] "Y-FGCZCont00100" "Y-FGCZCont00147" "Y-FGCZCont00243"
 [4] "Y-FGCZCont00285" "Y-FGCZCont00056" "Y-FGCZCont00010"
 [7] "Y-FGCZCont00001" "Y-FGCZCont00027" "Y-FGCZCont00248"
[10] "Y-FGCZCont00482" "Y-FGCZCont00093" "Y-FGCZCont00160"
[13] "Y-FGCZCont00175" "Y-FGCZCont00142" "Y-FGCZCont00066"
[16] "Y-FGCZCont00165" "Y-FGCZCont00099" "Y-FGCZCont00190"
[19] "Y-FGCZCont00162" "Y-FGCZCont00278" "Y-FGCZCont00116"
[22] "Y-FGCZCont00106" "Y-FGCZCont00122" "Q6ZSR6"         
[25] "Q6ZSR9"          "Y-FGCZCont00065" "Y-FGCZCont00282"
[28] "Y-FGCZCont00030" "Y-FGCZCont00113" "Y-FGCZCont00118"
[31] "Y-FGCZCont00378" "Y-FGCZCont00124" "Y-FGCZCont00375"
[34] "Y-FGCZCont00168" "Y-FGCZCont00180" "Y-FGCZCont00256"
[37] "Y-FGCZCont00260" "Y-FGCZCont00380" "Y-FGCZCont00272"
[40] "Y-FGCZCont00293" "Y-FGCZCont00301" "Y-FGCZCont00307"
[43] "Y-FGCZCont00311" "Y-FGCZCont00319" "Y-FGCZCont00330"
[46] "Y-FGCZCont00343" "Y-FGCZCont00347" "Y-FGCZCont00348"
[49] "Y-FGCZCont00351" "Y-FGCZCont00356" "Y-FGCZCont00359"
[52] "Y-FGCZCont00373" "Y-FGCZCont00374" "Y-FGCZCont00461"
[55] "Y-FGCZCont00459" "Y-FGCZCont00463" "Y-FGCZCont00474"
[58] "Y-FGCZCont00212" "Y-FGCZCont00231" "Y-FGCZCont00376"
[61] "Y-FGCZCont00189" "Y-FGCZCont00276" "Y-FGCZCont00367"
[64] "Y-FGCZCont00328" "Y-FGCZCont00355" "Y-FGCZCont00111"
[67] "Y-FGCZCont00054" "Y-FGCZCont00310" "Y-FGCZCont00188"
[70] "Y-FGCZCont00288" "Y-FGCZCont00418" "Y-FGCZCont00417"
[73] "Y-FGCZCont00422" "Y-FGCZCont00419" "Y-FGCZCont00151"
[76] "Y-FGCZCont00148" "Y-FGCZCont00033" "Y-FGCZCont00024"
[79] "Y-FGCZCont00035" "Y-FGCZCont00060" "Y-FGCZCont00237"
[82] "Y-FGCZCont00289" "Y-FGCZCont00098" "Y-FGCZCont00032"
[85] "Y-FGCZCont00329" "Y-FGCZCont00193" "Y-FGCZCont00150"
[88] "Y-FGCZCont00195" "Y-FGCZCont00414" "Y-FGCZCont00064"
[91] "Y-FGCZCont00110" "Y-FGCZCont00037" "Y-FGCZCont00251"
[94] "Y-FGCZCont00077" "Y-FGCZCont00127" "Y-FGCZCont00161"
[97] "Y-FGCZCont00286" "Y-FGCZCont00052"
#they can be safely removed
idTab.miss <- filter(idTab.miss, !is.na(hgnc_symbol)) %>%
  dplyr::select(-ensembl_gene_id)

Get ensemble IDs for those proteins using symbol

ids <- idTab.miss$hgnc_symbol
anno <- getBM(attributes=c('hgnc_symbol','ensembl_gene_id'), 
      filters = 'hgnc_symbol', 
      values = ids, 
      mart = ensembl)
idTab.miss <- left_join(idTab.miss, anno, by = "hgnc_symbol")

Genes whose ensembl ID still can not found, use manual annotation

mapSE <- structure(c("ENSG00000180389","ENSG00000282100","ENSG00000180448",
                     NA,NA,NA,"ENSG00000177144","ENSG00000223614","ENSG00000160221",
                     "ENSG00000280071","ENSG00000278615","ENSG00000282651",
                    "ENSG00000275895"),
  names=c("ATP5F1EP2","HSP90AB4P","ARHGAP45","IGHV3-43D","GAGE7","IGHV3-30-3","NUDT4B",
          "ZNF735","GATD3A","GATD3B","C11orf98","IGHV5-10-1","U2AF1L5"))

idTab.miss <- mutate(idTab.miss, ensembl_gene_id = ifelse(is.na(ensembl_gene_id), mapSE[hgnc_symbol], ensembl_gene_id))

Update annotations for proteins with missing annotation

idTab <- bind_rows(filter(idTab, !is.na(ensembl_gene_id)), idTab.miss)

Retrieve chromosome information (using ensembl id)

#firstly based on id
ids <- idTab$ensembl_gene_id
anno <- getBM(attributes=c('ensembl_gene_id','chromosome_name'), 
      filters = 'ensembl_gene_id', 
      values = ids,
      mart = ensembl)

idTab <- mutate(idTab, 
                chromosome_name = anno[match(idTab$ensembl_gene_id,anno$ensembl_gene_id),]$chromosome_name) %>%
  filter(!grepl("CHR|HG",chromosome_name)) %>%
  arrange(ensembl_gene_id) %>%
  distinct(uniprotID, .keep_all = TRUE)  #some proteins can be mapped to several genes, this is normal and mostly happens to histones. This step will remove then and only keep one. This should be fine. 


filter(idTab, is.na(chromosome_name))
# A tibble: 7 x 5
  ID                  uniprotID hgnc_symbol ensembl_gene_id chromosome_name
  <chr>               <chr>     <chr>       <chr>           <chr>          
1 P0DN76;Q01081       P0DN76    U2AF1L5     ENSG00000275895 <NA>           
2 E9PRG8              E9PRG8    C11orf98    ENSG00000278615 <NA>           
3 A0A0B4J2D5;P0DPI2   A0A0B4J2… GATD3B      ENSG00000280071 <NA>           
4 Q58FF6              Q58FF6    HSP90AB4P   ENSG00000282100 <NA>           
5 A0A0C4DH38;A0A0J9Y… A0A0J9YX… IGHV5-10-1  ENSG00000282651 <NA>           
6 P01782;P0DP04       P0DP04    IGHV3-43D   <NA>            <NA>           
7 P0DP02              P0DP02    IGHV3-30-3  <NA>            <NA>           

For proteins chromosome can not be annotated, manually add chromosome info

chrMap <- structure(c("15","21","11","14","14","21"),
                      names = c("Q58FF6","A0A0B4J2D5",
                                     "E9PRG8","A0A0J9YXX1",
                                     "P0DP04","P0DN76"))
idTab <- mutate(idTab, chromosome_name = ifelse(is.na(chromosome_name),
                                                chrMap[uniprotID],chromosome_name))

Annotate

rawTab <- sumToTiday(protCLL, rowID = "rowID", colID = "patID")
rawTab <- left_join(rawTab, idTab, by = c(name = "ID")) %>% 
  filter(!is.na(uniprotID)) %>% select(-rowID, -ID)

normTab <- sumToTiday(protCLL_norm, rowID = "rowID", colID = "patID")
normTab <- left_join(normTab, idTab, by = c(name = "ID")) %>% 
  filter(!is.na(uniprotID)) %>% select(-rowID, -ID)

In the LUMOS dataset, one protein identifier can be mapped to several proteins and genes. This may be because some peptides can not be uniquely mapped to a protein. In the analysis, I just duplicated the values for different proteins share the same identifier.

Combine and save objects

Combine table

protCLL_raw <- tidyToSum(rawTab, "uniprotID","patID", values = "count",
                         annoCol  = colnames(patAnno)[colnames(patAnno)!="patID"],
                         annoRow  = c("name","ensembl_gene_id","hgnc_symbol","chromosome_name"))

protCLL <-  tidyToSum(normTab, "uniprotID","patID", values = c("count", "QRILC"),
                         annoCol  = colnames(patAnno)[colnames(patAnno)!="patID"],
                         annoRow  = c("name","ensembl_gene_id","hgnc_symbol","chromosome_name"))

Annotate proteins that can not be uniquely mapped

#for all proteins
dupTab <- rowData(protCLL_raw) %>% data.frame(stringsAsFactors = FALSE) %>% rownames_to_column("id") %>%
  group_by(name) %>% summarise(n=length(id)) %>% filter(n>1)
rowData(protCLL)$uniqueMap <- ! rowData(protCLL)$name %in% dupTab$name
rowData(protCLL_raw)$uniqueMap <- ! rowData(protCLL_raw)$name %in% dupTab$name

#a list of proteins that can not be uniquely mapped
unique(rowData(protCLL[!rowData(protCLL)$uniqueMap,])$hgnc_symbol)
  [1] "NUDT4B"     "GATD3B"     "POM121C"    "SNRPGP15"   "ZNF732"    
  [6] "GPR89A"     "WASH3P"     "NACA"       "TIMM23"     "MYL12B"    
 [11] "TOX3"       "RBFOX2"     "EIF1B"      "PHF2"       "UGT2B17"   
 [16] "PIAS1"      "PIAS2"      "TOX4"       "KRT6A"      "MT2A"      
 [21] "MT1E"       "HIST1H2AB"  "CDK1"       "PRSS1"      "H2AFZ"     
 [26] "HIST1H2AK"  "ZNF735"     "GPR89B"     "SULT1A3"    "SULT1A4"   
 [31] "HSPA1A"     "HSPA1B"     "U2AF1L5"    "IGLC2"      "IGLC3"     
 [36] "CALM1"      "CALM2"      "CALM3"      "GATD3A"     "SLC2A3"    
 [41] "MT1G"       "SNRPB"      "ZNF43"      "MYL12A"     "CDK2"      
 [46] "RBMS1"      "HIST1H2BB"  "EIF1"       "KRT6C"      "SULT1A1"   
 [51] "UBE2D1"     "UGT2B15"    "ATP5E"      "ACTB"       "ARF3"      
 [56] "SNRPG"      "ACTA2"      "RPL39"      "GNAS"       "SNRPN"     
 [61] "ACTG1"      "ACTG2"      "HIST1H3B"   "NOMO3"      "MT1X"      
 [66] "ARF1"       "H3F3B"      "CCZ1B"      "CCZ1"       "CDK3"      
 [71] "U2AF1"      "ZNF85"      "RAB31"      "GTF2H2"     "ZNF141"    
 [76] "HIST2H2AC"  "HIST2H2BE"  "ORMDL2"     "RPL39P5"    "ALG10"     
 [81] "ALG10B"     "NOMO2"      "HIST2H2BF"  "TIMM23B"    "ATP5EP2"   
 [86] "HIST2H2AA3" "LAIR1"      "LAIR2"      "SLC25A25"   "GTF2H2C"   
 [91] "WASH2P"     "RBMS3"      "HIST2H3D"   "H2AFV"      "HIST3H2A"  
 [96] "MOB1B"      "ZNF679"     "MT1M"       "LILRB2"     "LILRB1"    
[101] "PRSS3P2"    "SLC2A14"    "HIST1H2BH"  "SLC25A36"   "POM121"    
[106] "FEM1C"      "HIST1H2AH"  "CNOT6L"     "ZNF681"     "CLIC6"     
[111] "HIST1H2AJ"  "SLC25A33"   "FEM1A"      "H2AFJ"      "SLC25A23"  
[116] "TRIM34"     "PANK2"      "MOB1A"      "PANK3"      "SPINK5"    
[121] "RBFOX1"     "CLIC5"      "NUDT4"      "ORMDL1"     "RAB22A"    
[126] "CNOT6"      "PHF8"       "UBE2D4"    

Save object

#for other projects
save(protCLL, protCLL_raw, file = "../../var/proteomic_LUMOS_20200430.RData")

#for this project
save(protCLL, protCLL_raw, file = "../output/proteomic_LUMOS_20200430.RData")

```


sessionInfo()
R version 3.6.0 (2019-04-26)
Platform: x86_64-apple-darwin15.6.0 (64-bit)
Running under: macOS  10.15.4

Matrix products: default
BLAS:   /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRblas.0.dylib
LAPACK: /Library/Frameworks/R.framework/Versions/3.6/Resources/lib/libRlapack.dylib

locale:
[1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8

attached base packages:
[1] stats4    parallel  stats     graphics  grDevices utils     datasets 
[8] methods   base     

other attached packages:
 [1] forcats_0.4.0               stringr_1.4.0              
 [3] dplyr_0.8.5                 purrr_0.3.3                
 [5] readr_1.3.1                 tidyr_1.0.0                
 [7] tibble_3.0.0                tidyverse_1.3.0            
 [9] SummarizedExperiment_1.14.0 DelayedArray_0.10.0        
[11] BiocParallel_1.18.0         matrixStats_0.54.0         
[13] GenomicRanges_1.36.0        GenomeInfoDb_1.20.0        
[15] IRanges_2.18.1              S4Vectors_0.22.0           
[17] biomaRt_2.40.0              DEP_1.6.1                  
[19] jyluMisc_0.1.5              vsn_3.52.0                 
[21] Biobase_2.44.0              BiocGenerics_0.30.0        
[23] pheatmap_1.0.12             cowplot_0.9.4              
[25] ggplot2_3.3.0               limma_3.40.2               

loaded via a namespace (and not attached):
  [1] utf8_1.1.4             shinydashboard_0.7.1   gmm_1.6-2             
  [4] tidyselect_1.0.0       RSQLite_2.1.1          AnnotationDbi_1.46.0  
  [7] htmlwidgets_1.3        grid_3.6.0             norm_1.0-9.5          
 [10] maxstat_0.7-25         munsell_0.5.0          codetools_0.2-16      
 [13] preprocessCore_1.46.0  DT_0.7                 withr_2.1.2           
 [16] colorspace_1.4-1       knitr_1.23             rstudioapi_0.10       
 [19] ggsignif_0.5.0         mzID_1.22.0            labeling_0.3          
 [22] git2r_0.26.1           slam_0.1-45            GenomeInfoDbData_1.2.1
 [25] KMsurv_0.1-5           farver_2.0.3           bit64_0.9-7           
 [28] rprojroot_1.3-2        vctrs_0.2.4            generics_0.0.2        
 [31] TH.data_1.0-10         xfun_0.8               sets_1.0-18           
 [34] R6_2.4.0               doParallel_1.0.14      clue_0.3-57           
 [37] bitops_1.0-6           fgsea_1.10.0           assertthat_0.2.1      
 [40] promises_1.0.1         scales_1.1.0           multcomp_1.4-10       
 [43] gtable_0.3.0           affy_1.62.0            sandwich_2.5-1        
 [46] workflowr_1.6.0        rlang_0.4.5            cmprsk_2.2-8          
 [49] mzR_2.18.1             GlobalOptions_0.1.0    splines_3.6.0         
 [52] impute_1.58.0          hexbin_1.27.3          broom_0.5.2           
 [55] modelr_0.1.5           BiocManager_1.30.4     yaml_2.2.0            
 [58] abind_1.4-5            backports_1.1.4        httpuv_1.5.1          
 [61] tools_3.6.0            relations_0.6-8        affyio_1.54.0         
 [64] ellipsis_0.2.0         gplots_3.0.1.1         RColorBrewer_1.1-2    
 [67] MSnbase_2.10.1         Rcpp_1.0.1             plyr_1.8.4            
 [70] progress_1.2.2         visNetwork_2.0.7       zlibbioc_1.30.0       
 [73] RCurl_1.95-4.12        prettyunits_1.0.2      ggpubr_0.2.1          
 [76] GetoptLong_0.1.7       zoo_1.8-6              haven_2.2.0           
 [79] cluster_2.1.0          exactRankTests_0.8-30  fs_1.4.0              
 [82] magrittr_1.5           data.table_1.12.2      openxlsx_4.1.0.1      
 [85] circlize_0.4.6         reprex_0.3.0           survminer_0.4.4       
 [88] pcaMethods_1.76.0      mvtnorm_1.0-11         whisker_0.3-2         
 [91] ProtGenerics_1.16.0    hms_0.5.2              shinyjs_1.0           
 [94] mime_0.7               evaluate_0.14          xtable_1.8-4          
 [97] XML_3.98-1.20          rio_0.5.16             readxl_1.3.1          
[100] gridExtra_2.3          shape_1.4.4            compiler_3.6.0        
[103] KernSmooth_2.23-15     ncdf4_1.16.1           crayon_1.3.4          
[106] htmltools_0.4.0        mgcv_1.8-28            later_0.8.0           
[109] lubridate_1.7.4        DBI_1.0.0              dbplyr_1.4.2          
[112] ComplexHeatmap_2.0.0   MASS_7.3-51.4          tmvtnorm_1.4-10       
[115] Matrix_1.2-17          car_3.0-3              cli_1.1.0             
[118] imputeLCMD_2.0         marray_1.62.0          gdata_2.18.0          
[121] igraph_1.2.4.1         pkgconfig_2.0.2        km.ci_0.5-2           
[124] foreign_0.8-71         piano_2.0.2            xml2_1.2.2            
[127] MALDIquant_1.19.3      foreach_1.4.4          XVector_0.24.0        
[130] drc_3.0-1              rvest_0.3.5            digest_0.6.19         
[133] rmarkdown_1.13         cellranger_1.1.0       fastmatch_1.1-0       
[136] survMisc_0.5.5         curl_3.3               shiny_1.3.2           
[139] gtools_3.8.1           rjson_0.2.20           lifecycle_0.2.0       
[142] nlme_3.1-140           jsonlite_1.6           carData_3.0-2         
[145] fansi_0.4.0            pillar_1.4.3           lattice_0.20-38       
[148] httr_1.4.1             plotrix_3.7-6          survival_2.44-1.1     
[151] glue_1.3.2             zip_2.0.2              png_0.1-7             
[154] iterators_1.0.10       bit_1.1-14             stringi_1.4.3         
[157] blob_1.1.1             memoise_1.1.0          caTools_1.17.1.2