Edward, born at Bisham (Berkshire) in 1560, was eldest son of Sir Thomas Hoby of Bisham Abbey (Berkshire), by Elizabeth the third daughter of Sir Anthony Cooke of Gidea Hall (Essex). He was educated at Eton, where he formed a lasting friendship with Sir John Harington. He matriculated at Oxford as a gentleman-commoner from Trinity College on 11th November 1574. He was allowed to graduate as a BA, on 19th February 1576, after keeping only eight terms and, before he had completed ten terms, proceeded to become an MA, on 3rd July of the same year, being the senior master in the comitia. At college, Thomas Lodge, the dramatist, was servitour or scholar under him. In June 1576, he obtained a dispensation for two years and two terms in order to travel on the continent. Subsequently, as he states in his 'Counter-snarle', he entered himself at the Middle Temple.

Under the auspices of his maternal uncle, Lord Burghley, Edward rose into high favour at court and was frequently employed on confidential missions. His fortunes were further advanced by his marriage, on 21st May 1582, with Margaret the daughter of Queen Elizabeth's cousin, Henry Carey, Lord Hunsdon. The day after the wedding, he was knighted by the Queen.

In August 1584, Sir Edward accompanied his father-in-law on a special mission to Scotland. His affability and learning greatly impressed King James VI and, after attending the Scottish Ambassador, Patrick, Master of Gray, as far as Durham, Hoby received a flattering letter from the Scottish king, dated 24th Octover 1584, in which James intimated his longing for his company, and how he had "commanded his ambassador to sue for it." Arran also wrote to the same effect and enclosed a "small token," which he begged Hoby to wear in 'testimony of their brotherhood. These amenities proved displeasing to Queen Elizabeth and Hoby found it convenient, for a time, to plead the ague as an excuse for not attending the court. Domestic troubles also harassed him.

On 24th September 1586, Sir Edward was returned as MP for Queenborough in Kent and gained distinction as a speaker in Parliament. On 31st October following, he complained that he had been "not only bitten but overpassed by the hard hand of Walsingham," and appealed to Secretary Davison to use his influence with the Queen on his behalf. Being ultimately restored to favour, Hoby, in July 1588, was chosen to report to the Queen the progress of the preparations against the Armada. In the ensuing October, he was elected MP for Berkshire and he was made JP for Middlesex by a special renewal of the commission on 17th December 1591. In 1592, the Queen visited Sir Edward and his mother at Bisham Abbey. He was chosen MP for Kent in February 1593 and, in 1594, was granted letters patent for buying and providing wool "for sale in England for ten years" and the grant was ratified in the succeeding reign.

Hoby accompanied the expedition to Cadiz in 1596, was made Constable of Queenborough Castle on the Isle of Sheppey (Kent), on 9th July 1597, and, on the following 28th October, received a commission to search out and prosecute all offences against the statute prohibiting the exportation of iron from England. His reward being half the forfeitures arising therefrom. He represented Rochester in the parliaments of 1597, 1601, February 1604 and 1614. James I made Sir Edward a gentleman of the Privy Chamber, forgave him, by warrant dated 7th January 1605, for the arrears of rent of the Royal Manor of Shirland in Derbyshire - amounting to over £500 - and, on 21st August 1607, granted him an exclusive license to buy wool in Warwickshire and Staffordshire. He frequently entertained the King at Bisham.

Hoby married at least three times and died at Queenborough Castle on 1st March 1617. He was buried in Bisham Church. By his first wife, who died in 1605, he had no issue, but he left an illegitimate son by his lover, Catherine Pinkney, named Peregrine (1602-1678) whom he brought up and made his heir. Upon Sir Edward's death, the boy's care was committed to Archbishop Abbot. Peregrine eventually sat in Parliament for Great Marlow in Buckinghamshire, in 1640, 1660 and 1661; and, in 1666, his eldest son, Edward, was, created a baronet.

An excellent scholar himself, Hoby cultivated the friendship of learned men, especially that of William Camden, who eulogises his bounty and accomplishments in his "Britannia" (under 'Bisham' and 'Queenborough'). Camden also dedicated his 'Hibernia' (1587) to Sir Edward. In 1612, Hoby presented Sir Henry Savile's sumptuous edition of 'St. Chrysostom' to the library of Trinity College, Oxford. Hoby was also a keen theologian, as his contests with the Papists, Theophilus Higgons and John Floyd sufficiently prove. He wrote several works, including 'A Letter to Mr. T[heophilus] H[iggons], late Minister: now Fugitive . . . in answer of his first Motive' (1609), which was answered by Higgons during the same year; 'A Counter-snarle for Ishmael Rabshacheh, a Cecropidan Lycaonite' (1613), being a reply to 'The Overthrow of the Protestants Pulpet Babels' by John Floyd, after which Floyd rejoined with his 'Purgatories triumph over Hell, maugre the barking of Cerberus in Sir Edward Hoby's Counter-snarle' (1613); and 'A Curry-combe for a coxe-combe....In answer to a lewd Libel lately foricated by Jabal Rachil against Sir Edward Hoby's Counter-Snarle, entitled Purgatories triumph over Hell' (1615), written under the ponderous pseudonym of 'Nick Groome of the Hoby Stable Regino-burgi' in the form of a dialogue. Sir Edward also translated, from the French of M. Coignet, 'Politique [discourses on] truth and lying' (1586); and, from the Spanish of B. de Meudoza, 'Theorique and Practise of Warre,' (1597).